Compounds as rearranged during transfection (RET) inhibitors

ABSTRACT

This invention relates to novel compounds which are inhibitors of the Rearranged during Transfection (RET) kinase, to pharmaceutical compositions containing them, to processes for their preparation, and to their use in therapy, alone or in combination, for the normalization of gastrointestinal sensitivity, motility and/or secretion and/or abdominal disorders or diseases and/or treatment related to diseases related to RET dysfunction or where modulation of RET activity may have therapeutic benefit including but not limited to all classifications of irritable bowel syndrome (IBS) including diarrhea-predominant, constipation-predominant or alternating stool pattern, functional bloating, functional constipation, functional diarrhea, unspecified functional bowel disorder, functional abdominal pain syndrome, chronic idiopathic constipation, functional esophageal disorders, functional gastroduodenal disorders, functional anorectal pain, inflammatory bowel disease, proliferative diseases such as non-small cell lung cancer, hepatocellular carcinoma, colorectal cancer, medullary thyroid cancer, follicular thyroid cancer, anaplastic thyroid cancer, papillary thyroid cancer, brain tumors, peritoneal cavity cancer, solid tumors, other lung cancer, head and neck cancer, gliomas, neuroblastomas, Von Hippel-Lindau Syndrome and kidney tumors, breast cancer, fallopian tube cancer, ovarian cancer, transitional cell cancer, prostate cancer, cancer of the esophagus and gastroesophageal junction, biliary cancer, adenocarcinoma, and any malignancy with increased RET kinase activity.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/145,271, filed Sep. 28, 2018, which is a division of Ser. No.15/877,524, filed Jan. 23, 2018, which is a continuation of Ser. No.15/509,613, filed Mar. 8, 2017, which is a 371 National Stage Entry ofInternational Application No. PCT/M2015/056905, filed Sep. 9, 2015,which claim priorities to International Application Nos.PCT/CN2015/086995, filed Aug. 14, 2015, and PCT/CN2014/086 197, filedSep. 10, 2014.

FIELD OF INVENTION

This invention relates to novel compounds which are inhibitors of theRearranged during Transfection (RET) kinase, to pharmaceuticalcompositions containing them, to processes for their preparation, and totheir use in therapy, alone or in combination, for the normalization ofgastrointestinal sensitivity, motility and/or secretion and/or abdominaldisorders or diseases and/or treatment related to diseases related toRET dysfunction or where modulation of RET activity may have therapeuticbenefit including but not limited to all classifications of irritablebowel syndrome (IBS) including diarrhea-predominant,constipation-predominant or alternating stool pattern, functionalbloating, functional constipation, functional diarrhea, unspecifiedfunctional bowel disorder, functional abdominal pain syndrome, chronicidiopathic constipation, functional esophageal disorders, functionalgastroduodenal disorders, functional anorectal pain, inflammatory boweldisease, proliferative diseases such as non-small cell lung cancer,hepatocellular carcinoma, colorectal cancer, medullary thyroid cancer,follicular thyroid cancer, anaplastic thyroid cancer, papillary thyroidcancer, brain tumors, peritoneal cavity cancer, solid tumors, other lungcancer, head and neck cancer, gliomas, neuroblastomas, Von Hippel-LindauSyndrome and kidney tumors, breast cancer, fallopian tube cancer,ovarian cancer, transitional cell cancer, prostate cancer, cancer of theesophagus and gastroesophageal junction, biliary cancer andadenocarcinoma, and any malignancy with increased RET kinase activity.

BACKGROUND OF THE INVENTION

Irritable bowel syndrome (IBS) is a common illness affecting 10-20% ofindividuals in developed countries and is characterized by abnormalbowel habits, bloating and visceral hypersensitivity (Camilleri, M., N.Engl. J. Med., 2012, 367:1626-1635). While the etiology of IBS isunknown it is thought to result from either a disorder between the brainand gastrointestinal tract, a disturbance in the gut microbiome orincreased inflammation. The resulting gastrointestinal changes affectnormal bowel transit resulting in either diarrhea or constipation.Furthermore in a majority of IBS patients the sensitization of theperipheral nervous system results in visceral hypersensitivity orallodynia (Keszthelyi, D., Eur. J. Pain, 2012, 16:1444-1454).

While IBS does not directly alter life expectancy it has a considerableeffect on a patient's quality of life. Moreover there is a significantfinancial cost for IBS associated healthcare and lost productivity dueto worker absenteeism (Nellesen, D., et al., J. Manag. Care Pharm.,2013, 19:755-764). One of the most important symptoms that greatlyaffect an IBS patient's quality of life is visceral pain (Spiegel, B.,et al., Am. J. Gastroenterol., 2008, 103:2536-2543). Molecularstrategies that inhibit IBS associated visceral pain would greatlyinfluence the IBS patient's quality of life and reduce associated costs.

Rearranged during transfection (RET) is a neuronal growth factorreceptor tyrosine kinase that is activated upon binding one of fourneurotrophic factors glial cell line-derived neurotrophic factor (GDNF),neurturin, artemin and persephin in combination with a co-receptor GDNFfamily receptor alpha-1, 2, 3, and 4 respectively (Plaza-Menacho, I., etal., Trends Genet., 2006, 22:627-636). RET is known to play an importantrole in the development and survival of afferent nociceptors in the skinand gut. RET kinase knock-out mice lack enteric neurons and have othernervous system anomalies suggesting that a functional RET kinase proteinproduct is required during development (Taraviras, S. et al.,Development, 1999, 126:2785-2797). Moreover population studies ofpatients with Hirschsprung's disease characterized by colonicobstruction due to lack of normal colonic enervation have a higherproportion of both familial and sporadic loss of function RET mutations(Butler Tjaden N., et al., Transl. Res., 2013, 162:1-15).

Similarly, aberrant RET kinase activity is associated with multipleendocrine neoplasia (MEN 2A and 2B), familial medullary thyroidcarcinoma (FMTC), papillary thyroid carcinoma (PTC) and Hirschsprung'sdisease (HSCR) (Borello, M., et al., Expert Opin. Ther. Targets, 2013,17:403-419). MEN 2A is a cancer syndrome resulting from a mutation inthe extracellular cysteine-rich domain of RET leading to dimerizationvia a disulfide bond which causes constitutive activation of thetyrosine kinase activity (Wells Jr, S., et al., J. Clin. Endocrinol.Metab., 2013, 98:3149-3164). Individuals with this mutation may developmedullary thyroid carcinoma (MTC), parathyroid hyperplasia, andpheochromocytoma. MEN 2B is caused by a Met918Thr mutation in RET whichchanges the tyrosine kinase specificity. MEN 2B is similar to MEN 2A,but lacks the parathyroid hyperplasia and also leads to development ofnumerous mucosal ganglia of the lips, tongue, and intestinal tract.Chromosomal rearrangements linking the promoter and NH2-terminal domainsor unrelated gene(s) to the COOH-terminus of RET kinase resulting inconstitutively activated chimeric forms of the receptor (RET/PTC) arethought to be tumor initiating events in PTC (Viglietto, G. et al.,Oncogene, 1995, 11:1207-1210). PTC's encompass about 80% of all thyroidcarcinomas. These data indicate that inhibition of RET may be anattractive therapeutic strategy for the treatment of pain associatedwith IBS and other gastrointestinal disorders and for the treatment ofcancers with constitutive RET kinase activity.

SUMMARY OF THE INVENTION

This invention relates toN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide,represented by Formula (I):

or pharmaceutically acceptable salts thereof, and crystalline formsthereof, and toN-(3-(2-(dimethylamino)ethoxy)-5-(trifluoromethyl)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide,represented by Formula (II):

or pharmaceutically acceptable salts thereof.

This invention also relates to a pharmaceutical composition comprising acompound of Formula (I) or (II) and a pharmaceutically acceptableexcipient.

This invention also relates to a method of treating irritable bowelsyndrome comprising administering to a human in need thereof aneffective amount of a compound of Formula (I) or (II) or apharmaceutically acceptable salt thereof. This invention also relates toa method of treating cancer comprising administering to a human in needthereof an effective amount of a compound of Formula (I) or (II) or apharmaceutically acceptable salt thereof.

This invention also relates to compounds of Formula (I) or (II) for usein therapy. This invention also relates to the use of a compound ofFormula (I) or (II) or a pharmaceutically acceptable salt thereof forthe treatment of irritable bowel syndrome. This invention also relatesto the use of a compound of Formula (I) or (II) or a pharmaceuticallyacceptable salt thereof for the treatment of cancer.

This invention also relates to the use of a compound of Formula (I) or(II) or a pharmaceutically acceptable salt thereof in the preparation ofa medicament for the treatment of diseases mediated by RET. Thisinvention also relates to the use of a compound of Formula (I) or (II)or a pharmaceutically acceptable salt thereof in the manufacture of amedicament for the treatment of irritable bowel syndrome. This inventionalso relates to the use of a compound of Formula (I) or (II) or apharmaceutically acceptable salt thereof in the manufacture of amedicament for the treatment of cancer.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows an X-ray powder diffraction pattern of hydrate 1 of thehydrochloric acid salt of the compound of Formula (I).

FIG. 2 shows a Raman spectrum of hydrate 1 of the hydrochloric acid saltof the compound of Formula (I).

FIG. 3 shows a differential scanning calorimetry trace of hydrate 1 ofthe hydrochloric acid salt of the compound of Formula (I).

FIG. 4 shows a thermogravimetric analysis trace of hydrate 1 of thehydrochloric acid salt of the compound of Formula (I).

FIG. 5 shows an X-ray powder diffraction pattern of hydrate 2 of thehydrochloric acid salt of the compound of Formula (I).

FIG. 6 shows a Raman spectrum of hydrate 2 of the hydrochloric acid saltof the compound of Formula (I).

FIG. 7 shows a differential scanning calorimetry trace of hydrate 2 ofthe hydrochloric acid salt of the compound of Formula (I).

FIG. 8 shows a thermogravimetric analysis trace of hydrate 2 of thehydrochloric acid salt of the compound of Formula (I).

FIG. 9 shows an X-ray powder diffraction pattern of the anhydroushydrochloric acid salt of the compound of Formula (I).

FIG. 10 shows a Raman spectrum of the anhydrate of the anhydroushydrochloric acid salt of the compound of Formula (I).

FIG. 11 shows a differential scanning calorimetry trace of the anhydroushydrochloric acid salt of the compound of Formula (I).

FIG. 12 shows a thermogravimetric analysis trace of the anhydroushydrochloric acid salt of the compound of Formula (I).

FIG. 13 shows an X-ray powder diffraction pattern of the aspartic acidsalt of the compound of Formula (I).

FIG. 14 shows a Raman spectrum of the aspartic acid salt of the compoundof Formula (I).

FIG. 15 shows a differential scanning calorimetry trace of the asparticacid salt of the compound of Formula (I).

FIG. 16 shows a thermogravimetric analysis trace of the aspartic acidsalt of the compound of Formula (I).

FIG. 17 shows an X-ray powder diffraction pattern of the hippuric acidsalt of the compound of Formula (I).

FIG. 18 shows a Raman spectrum of the hippuric acid salt of the compoundof Formula (I).

FIG. 19 shows a differential scanning calorimetry trace of the hippuricacid salt of the compound of Formula (I).

FIG. 20 shows a thermogravimetric analysis trace of the hippuric acidsalt of the compound of Formula (I).

FIG. 21 shows an X-ray powder diffraction pattern of the phophoric acidsalt of the compound of Formula (I).

FIG. 22 shows a Raman spectrum of the phophoric acid salt of thecompound of Formula (I).

FIG. 23 shows a differential scanning calorimetry trace of the phophoricacid salt of the compound of Formula (I).

FIG. 24 shows a thermogravimetric analysis trace of the phophoric acidsalt of the compound of Formula (I).

DETAILED DESCRIPTION OF THE INVENTION

This invention relates to compounds of the Formula (I) or (II) orpharmaceutically acceptable salts thereof as defined above.

A person of ordinary skills in the art recognizes that compounds of thepresent invention may have alternative names when different namingsoftware is used.

This invention also relates to compounds of Formula (I) or (II), orpharmaceutically acceptable salts thereof, for use in therapy, inparticular, for use in therapy wherein the subject is a human. Inparticular, for use in the treatment of diseases mediated by RET:irritable bowel syndrome (IBS) including diarrhea-predominant,constipation-predominant or alternating stool pattern, functionalbloating, functional constipation, functional diarrhea, unspecifiedfunctional bowel disorder, functional abdominal pain syndrome, chronicidiopathic constipation, functional esophageal disorders, functionalgastroduodenal disorders, functional anorectal pain, inflammatory boweldisease, proliferative diseases such as non-small cell lung cancer,hepatocellular carcinoma, colorectal cancer, medullary thyroid cancer,follicular thyroid cancer, anaplastic thyroid cancer, papillary thyroidcancer, brain tumors, peritoneal cavity cancer, solid tumors, other lungcancer, head and neck cancer, gliomas, neuroblastomas, Von Hippel-LindauSyndrome and kidney tumors, breast cancer, fallopian tube cancer,ovarian cancer, transitional cell cancer, prostate cancer, caner of theesophagus and gastroesophageal junction, biliary cancer andadenocarcinoma. In particular, this invention relates to compounds ofFormula (I) or (II), or pharmaceutically acceptable salts thereof, foruse in the treatment of irritable bowel syndrome (IBS) includingdiarrhea-predominant, constipation-predominant or alternating stoolpattern, functional bloating, functional constipation, functionaldiarrhea, unspecified functional bowel disorder, functional abdominalpain syndrome, chronic idiopathic constipation, functional esophagealdisorders, functional gastroduodenal disorders, functional anorectalpain, inflammatory bowel disease, non-small cell lung cancer,hepatocellular carcinoma, colorectal cancer, medullary thyroid cancer,follicular thyroid cancer, anaplastic thyroid cancer, papillary thyroidcancer, brain tumors, peritoneal cavity cancer, solid tumors, other lungcancer, head and neck cancer, gliomas, neuroblastomas, Von Hippel-LindauSyndrome and kidney tumors, breast cancer, fallopian tube cancer,ovarian cancer, transitional cell cancer, prostate cancer, cancer of theesophagus and gastroesophageal junction, biliary cancer andadenocarcinoma.

This invention also relates to compounds of Formula (I) or (II), orpharmaceutically acceptable salts thereof, for use as a medicament. Inanother embodiment, the invention relates to the use of compounds of theinvention in the preparation of a medicament for the treatment ofdiseases mediated by RET. This invention also relates to compounds ofFormula (I) or (II), or pharmaceutically acceptable salts thereof, inthe manufacture of a medicament for the treatment of irritable bowelsyndrome. This invention also relates to compounds of Formula (I) or(II), or pharmaceutically acceptable salts thereof, in the manufactureof a medicament for the treatment of cancer.

This invention also relates to the use of compounds of Formula (I) or(II) in therapy. The invention further includes the use of compounds ofthe invention as an active therapeutic substance, in particular in thetreatment of diseases mediated by RET. This invention also relates tothe use of compounds of Formula (I) or (II) for the treatment ofirritable bowel syndrome. This invention also relates to the use ofcompounds of Formula (I) or (II) for the treatment of cancer.

Because of their potential use in medicine, the salts of the compoundsof Formula (I) are preferably pharmaceutically acceptable. Suitablepharmaceutically acceptable salts include those described by Berge,Bighley, and Monkhouse, J. Pharm. Sci. (1977) 66, pp 1-19. Saltsencompassed within the term “pharmaceutically acceptable salts” refer tonon-toxic salts of the compounds of this invention. Salts of thedisclosed compounds may be prepared by any suitable method known in theart, including treatment of the free base with an inorganic acid, suchas hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like, or with an organic acid, such as aceticacid, trifluoroacetic acid, maleic acid, succinic acid, mandelic acid,fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid,salicylic acid, pyranosidyl acid, such as glucuronic acid orgalacturonic acid, alpha-hydroxy acid, such as citric acid or tartaricacid, amino acid, such as aspartic acid or glutamic acid, aromatic acid,such as benzoic acid or cinnamic acid, sulfonic acid, such asp-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid or thelike. Examples of pharmaceutically acceptable salts include sulfates,pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, chlorides,bromides, iodides, acetates, propionates, decanoates, caprylates,acrylates, formates, isobutyrates, caproates, heptanoates, propiolates,oxalates, malonates succinates, suberates, sebacates, fumarates,maleates, butyne-1,4-dioates, hexyne-1,6-dioates, benzoates,chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates,methoxybenzoates, phthalates, phenylacetates, phenylpropionates,phenylbutrates, citrates, lactates, γ-hydroxybutyrates, glycolates,tartrates mandelates, and sulfonates, such as xylenesulfonates,methanesulfonates, propanesulfonates, naphthalene-1-sulfonates andnaphthalene-2-sulfonates.

Pharmaceutically acceptable salt may also be made with a base whichaffords a pharmaceutically acceptable cation, which includes alkalimetal salts (especially sodium and potassium), alkaline earth metalsalts (especially calcium and magnesium), aluminum salts and ammoniumsalts, as well as salts made from physiologically acceptable organicbases such as trimethylamine, triethylamine, morpholine, pyridine,piperidine, picoline, dicyclohexylamine, N,N′-dibenzylethylenediamine,2-hydroxyethylamine, bis-(2-hydroxyethyl)amine,tri-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine,dehydroabietylamine, N,N′-bisdehydroabietylamine, glucamine,N-methylglucamine, collidine, choline, quinine, quinoline, and basicamino acid such as lysine and arginine.

Other salts, which are not pharmaceutically acceptable, may be useful inthe preparation of compounds of this invention and these should beconsidered to form a further aspect of the invention. These salts, suchas trifluoroacetate, while not in themselves pharmaceuticallyacceptable, may be useful in the preparation of salts useful asintermediates in obtaining the compounds of the invention and theirpharmaceutically acceptable salts.

If a compound of the invention is isolated as a salt, the correspondingfree base form of that compound may be prepared by any suitable methodknown to the art, including treatment of the salt with an inorganic ororganic base, suitably an inorganic or organic base having a higherpK_(a) than the free base form of the compound. Similarly, if a compoundof the invention is isolated as a salt, the corresponding free acid formof that compound may be prepared by any suitable method known to theart, including treatment of the salt with an inorganic or organic acid,suitably an inorganic or organic acid having a lower pK_(a) than thefree acid form of the compound.

The compound of Formula (I) or (II) may exist in a crystalline ornoncrystalline form, or as a mixture thereof. The skilled artisan willappreciate that pharmaceutically acceptable solvates may be formed forcrystalline or non-crystalline compounds. In crystalline solvates,solvent molecules are incorporated into the crystalline lattice duringcrystallization. Solvates may involve non-aqueous solvents such as, butnot limited to, ethanol, isopropanol, DMSO, acetic acid, ethanolamine,or ethyl acetate, or they may involve water as the solvent that isincorporated into the crystalline lattice. Solvates wherein water is thesolvent incorporated into the crystalline lattice are typically referredto as “hydrates.” Hydrates include stoichiometric hydrates as well ascompositions containing variable amounts of water. The inventionincludes all such solvates.

The skilled artisan will further appreciate that the compounds of theinvention that exist in crystalline form, including the various solvatesthereof, may exhibit polymorphism (i.e. the capacity to occur indifferent crystalline structures). These different crystalline forms aretypically known as “polymorphs.” The invention includes all suchpolymorphs. Polymorphs have the same chemical composition but differ inpacking, geometrical arrangement, and other descriptive properties ofthe crystalline solid state. Polymorphs, therefore, may have differentphysical properties such as shape, density, hardness, deformability,stability, and dissolution properties. Polymorphs typically exhibitdifferent melting points, IR spectra, and X-ray powder diffractionpatterns, which may be used for identification. The skilled artisan willappreciate that different polymorphs may be produced, for example, bychanging or adjusting the reaction conditions or reagents, used inmaking the compound. For example, changes in temperature, pressure, orsolvent may result in polymorphs. In addition, one polymorph mayspontaneously convert to another polymorph under certain conditions.

The present invention is further directed to certain crystalline formsof various salts ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide,in particular the hydrochloric acid salt, aspartic acid salt, hippuricacid salt, and phosphoric acid salt.

In some embodiments, a crystalline form ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride (hydrate 1 of the hydrochloric acid salt of the compoundof Formula (I)) is characterized by an X-ray powder diffraction (XRPD)pattern comprising at least nine diffraction angles, when measured usingCu K_(α) radiation, selected from a group consisting of about 6.0, 6.1,9.0, 9.2, 11.8, 11.9, 12.1, 13.3, 13.4, 13.6, 14.0, 14.8, 14.9, 15.3,20.5, 22.2, 22.3, 24.5, 24.6, 25.0, 25.1, and 25.2 degrees 2θ. Inanother embodiment, hydrate 1 of the hydrochloric acid salt of thecompound of Formula (I) is characterized by an X-ray powder diffraction(XRPD) pattern comprising at least eight diffraction angles or at leastseven diffraction angles or at least six diffraction angles or at leastfive diffraction angles or at least four diffraction angles, whenmeasured using Cu K_(α) radiation, selected from a group consisting ofabout 6.0, 6.1, 9.0, 9.2, 11.8, 11.9, 12.1, 13.3, 13.4, 13.6, 14.0,14.8, 14.9, 15.3, 20.5, 22.2, 22.3, 24.5, 24.6, 25.0, 25.1, and 25.2degrees 2θ. In another embodiment, hydrate 1 of the hydrochloric acidsalt of the compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern comprising at least three diffraction angles,when measured using Cu K_(α) radiation, selected from a group consistingof about 6.0, 6.1, 9.0, 9.2, 11.8, 11.9, 12.1, 13.3, 13.4, 13.6, 14.0,14.8, 14.9, 15.3, 20.5, 22.2, 22.3, 24.5, 24.6, 25.0, 25.1, and 25.2degrees 2θ.

In still another embodiment, hydrate 1 of the hydrochloric acid salt ofthe compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern comprising diffraction angles, when measuredusing Cu K_(α) radiation, of about 6.0, 9.0, 11.8, 12.1, 13.4, 14.8, and20.5 degrees 2θ. In yet another embodiment, hydrate 1 of thehydrochloric acid salt of the compound of Formula (I) is characterizedby an X-ray powder diffraction (XRPD) pattern substantially inaccordance with FIG. 1.

In other embodiments, hydrate 1 of the hydrochloric acid salt of thecompound of Formula (I) is characterized by a Raman spectrum comprisingat least nine peaks at positions selected from a group consisting ofpeaks at about 456, 581, 698, 774, 809, 952, 999, 1030, 1109, 1172,1248, 1282, 1335, 1362, 1461, 1532, 1626, 1678, 2903, 2952, and 3033cm⁻¹. In another embodiment, hydrate 1 of the hydrochloric acid salt ofthe compound of Formula (I) is characterized by a Raman spectrumcomprising at least eight peaks or at least seven peaks or at least sixpeaks or at least five peaks or at least four three peaks at positionsselected from a group consisting of peaks at about 456, 581, 698, 774,809, 952, 999, 1030, 1109, 1172, 1248, 1282, 1335, 1362, 1461, 1532,1626, 1678, 2903, 2952, and 3033 cm⁻¹. In another embodiment, hydrate 1of the hydrochloric acid salt of the compound of Formula (I) ischaracterized by a Raman spectrum comprising at least three peaks atpositions selected from a group consisting of peaks at about 456, 581,698, 774, 809, 952, 999, 1030, 1109, 1172, 1248, 1282, 1335, 1362, 1461,1532, 1626, 1678, 2903, 2952, and 3033 cm⁻¹.

In still another embodiment, hydrate 1 of the hydrochloric acid salt ofthe compound of Formula (I) is characterized by a Raman spectrumcomprising peaks at about 774, 809, 999, 1282, 1335, 1362, 1532, 1626,2903, 2952, and 3033 cm⁻¹. In yet another embodiment, hydrate 1 of thehydrochloric acid salt of the compound of Formula (I) is characterizedby a Raman spectrum substantially in accordance with FIG. 2.

In further embodiments, hydrate 1 of the hydrochloric acid salt of thecompound of Formula (I) is characterized by a differential scanningcalorimetry trace substantially in accordance with FIG. 3 and/or athermogravimetric analysis trace substantially in accordance with FIG.4.

In still further embodiments, as a person having ordinary skill in theart will understand, hydrate 1 of the hydrochloric acid salt of thecompound of Formula (I) is characterized by any combination of theanalytical data characterizing the aforementioned embodiments. Forexample, in one embodiment, hydrate 1 of the hydrochloric acid salt ofthe compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern substantially in accordance with FIG. 1 and aRaman spectrum substantially in accordance with FIG. 2 and adifferential scanning calorimetry trace substantially in accordance withFIG. 3 and a thermogravimetric analysis trace substantially inaccordance with FIG. 4. In another embodiment, hydrate 1 of thehydrochloric acid salt of the compound of Formula (I) is characterizedby an X-ray powder diffraction (XRPD) pattern substantially inaccordance with FIG. 1 and a Raman spectrum substantially in accordancewith FIG. 2. In another embodiment, hydrate 1 of the hydrochloric acidsalt of the compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern substantially in accordance with FIG. 1 and adifferential scanning calorimetry trace substantially in accordance withFIG. 3. In another embodiment, hydrate 1 of the hydrochloric acid saltof the compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern substantially in accordance with FIG. 1 and athermogravimetric analysis trace substantially in accordance with FIG.4. In another embodiment, hydrate 1 of the hydrochloric acid salt of thecompound of Formula (I) is characterized by an X-ray powder diffraction(XRPD) pattern comprising diffraction angles, when measured using CuK_(α) radiation, of about 6.0, 9.0, 11.8, 12.1, 13.4, 14.8, and 20.5degrees 2θ, and a Raman spectrum comprising peaks at about 774, 809,999, 1282, 1335, 1362, 1532, 1626, 2903, 2952, and 3033 cm⁻¹. In anotherembodiment, hydrate 1 of the hydrochloric acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising diffraction angles, when measured using Cu K_(α)radiation, of about 6.0, 9.0, 11.8, 12.1, 13.4, 14.8, and 20.5 degrees2θ, and a differential scanning calorimetry trace substantially inaccordance with FIG. 3. In another embodiment, hydrate 1 of thehydrochloric acid salt of the compound of Formula (I) is characterizedby an X-ray powder diffraction (XRPD) pattern comprising diffractionangles, when measured using Cu K_(α) radiation, of about 6.0, 9.0, 11.8,12.1, 13.4, 14.8, and 20.5 degrees 2θ, and a thermogravimetric analysistrace substantially in accordance with FIG. 4.

In some embodiments, a crystalline form ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride (hydrate 2 of the hydrochloric acid salt of the compoundof Formula (I)) is characterized by an X-ray powder diffraction (XRPD)pattern comprising at least nine diffraction angles, when measured usingCu K_(α) radiation, selected from a group consisting of about 6.0, 9.0,12.1, 13.4, 13.5, 14.7, 14.8, 14.9, 15.3, 20.3, 20.4, 22.2, 22.3, 22.4,24.7, and 24.8 degrees 2θ. In another embodiment, hydrate 2 of thehydrochloric acid salt of the compound of Formula (I) is characterizedby an X-ray powder diffraction (XRPD) pattern comprising at least eightdiffraction angles or at least seven diffraction angles or at least sixdiffraction angles or at least five diffraction angles or at least fourdiffraction angles, when measured using Cu K_(α) radiation, selectedfrom a group consisting of about 6.0, 9.0, 12.1, 13.4, 13.5, 14.7, 14.8,14.9, 15.3, 20.3, 20.4, 22.2, 22.3, 22.4, 24.7, and 24.8 degrees 2θ. Inanother embodiment, hydrate 2 of the hydrochloric acid salt of thecompound of Formula (I) is characterized by an X-ray powder diffraction(XRPD) pattern comprising at least three diffraction angles, whenmeasured using Cu K_(α) radiation, selected from a group consisting ofabout 6.0, 9.0, 12.1, 13.4, 13.5, 14.7, 14.8, 14.9, 15.3, 20.3, 20.4,22.2, 22.3, 22.4, 24.7, and 24.8 degrees 2θ.

In still another embodiment, hydrate 2 of the hydrochloric acid salt ofthe compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern comprising diffraction angles, when measuredusing Cu K_(α) radiation, of about 6.0, 9.0, 12.1, 14.7, 20.3, and 24.7degrees 2θ. In yet another embodiment, hydrate 2 of the hydrochloricacid salt of the compound of Formula (I) is characterized by an X-raypowder diffraction (XRPD) pattern substantially in accordance with FIG.5.

In other embodiments, hydrate 2 of the hydrochloric acid salt of thecompound of Formula (I) is characterized by a Raman spectrum comprisingat least nine peaks at positions selected from a group consisting ofpeaks at about 455, 588, 699, 734, 775, 807, 885, 949, 1000, 1033, 1112,1181, 1247, 1269, 1283, 1332, 1366, 1425, 1466, 1530, 1550, 1570, 1627,1684, 2902, 2946, and 3044 cm⁻¹. In another embodiment, hydrate 2 of thehydrochloric acid salt of the compound of Formula (I) is characterizedby a Raman spectrum comprising at least eight peaks or at least sevenpeaks or at least six peaks or at least five peaks or at least fourthree peaks at positions selected from a group consisting of peaks atabout 455, 588, 699, 734, 775, 807, 885, 949, 1000, 1033, 1112, 1181,1247, 1269, 1283, 1332, 1366, 1425, 1466, 1530, 1550, 1570, 1627, 1684,2902, 2946, and 3044 cm⁻¹. In another embodiment, hydrate 2 of thehydrochloric acid salt of the compound of Formula (I) is characterizedby a Raman spectrum comprising at least three peaks at positionsselected from a group consisting of peaks at about 455, 588, 699, 734,775, 807, 885, 949, 1000, 1033, 1112, 1181, 1247, 1269, 1283, 1332,1366, 1425, 1466, 1530, 1550, 1570, 1627, 1684, 2902, 2946, and 3044cm⁻¹.

In still another embodiment, hydrate 2 of the hydrochloric acid salt ofthe compound of Formula (I) is characterized by a Raman spectrumcomprising peaks at about 775, 1000, 1247, 1269, 1283, 1332, 1366, 1627,2902, and 2946 cm⁻¹. In yet another embodiment, hydrate 2 of thehydrochloric acid salt of the compound of Formula (I) is characterizedby a Raman spectrum substantially in accordance with FIG. 6.

In further embodiments, hydrate 2 of the hydrochloric acid salt of thecompound of Formula (I) is characterized by a differential scanningcalorimetry trace substantially in accordance with FIG. 7 and/or athermogravimetric analysis trace substantially in accordance with FIG.8.

In still further embodiments, as a person having ordinary skill in theart will understand, hydrate 2 of the hydrochloric acid salt of thecompound of Formula (I) is characterized by any combination of theanalytical data characterizing the aforementioned embodiments. Forexample, in one embodiment, hydrate 2 of the hydrochloric acid salt ofthe compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern substantially in accordance with FIG. 5 and aRaman spectrum substantially in accordance with FIG. 6 and adifferential scanning calorimetry trace substantially in accordance withFIG. 7 and a thermogravimetric analysis trace substantially inaccordance with FIG. 8. In another embodiment, hydrate 2 of thehydrochloric acid salt of the compound of Formula (I) is characterizedby an X-ray powder diffraction (XRPD) pattern substantially inaccordance with FIG. 5 and a Raman spectrum substantially in accordancewith FIG. 6. In another embodiment, hydrate 2 of the hydrochloric acidsalt of the compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern substantially in accordance with FIG. 5 and adifferential scanning calorimetry trace substantially in accordance withFIG. 7. In another embodiment, hydrate 2 of the hydrochloric acid saltof the compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern substantially in accordance with FIG. 5 and athermogravimetric analysis trace substantially in accordance with FIG.8. In another embodiment, hydrate 2 of the hydrochloric acid salt of thecompound of Formula (I) is characterized by an X-ray powder diffraction(XRPD) pattern comprising diffraction angles, when measured using CuK_(α) radiation, of about 6.0, 9.0, 12.1, 14.7, 20.3, and 24.7 degrees2θ, and a Raman spectrum comprising peaks at about 775, 1000, 1247,1269, 1283, 1332, 1366, 1627, 2902, and 2946 cm⁻¹. In anotherembodiment, hydrate 2 of the hydrochloric acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising diffraction angles, when measured using Cu K_(α)radiation, of about 6.0, 9.0, 12.1, 14.7, 20.3, and 24.7 degrees 2θ, anda differential scanning calorimetry trace substantially in accordancewith FIG. 7. In another embodiment, hydrate 2 of the hydrochloric acidsalt of the compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern comprising diffraction angles, when measuredusing Cu K_(α) radiation, of about 6.0, 9.0, 12.1, 14.7, 20.3, and 24.7degrees 2θ, and a thermogravimetric analysis trace substantially inaccordance with FIG. 8.

In some embodiments, a crystalline form ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride (the anhydrous hydrochloric acid salt of the compound ofFormula (I)) is characterized by an X-ray powder diffraction (XRPD)pattern comprising at least nine diffraction angles, when measured usingCu K_(α) radiation, selected from a group consisting of about 8.3, 8.4,10.7, 11.3, 15.5, 16.0, 20.0, 20.4, 20.8, 22.6, 23.2, 23.3, 23.6, 24.6,24.9, 25.3, 25.9, 26.9, 27.3, 27.4, 28.1, and 28.2 degrees 2θ. Inanother embodiment, the anhydrous hydrochloric acid salt of the compoundof Formula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising at least eight diffraction angles or at least sevendiffraction angles or at least six diffraction angles or at least fivediffraction angles or at least four diffraction angles, when measuredusing Cu K_(α) radiation, selected from a group consisting of about 8.3,8.4, 10.7, 11.3, 15.5, 16.0, 20.0, 20.4, 20.8, 22.6, 23.2, 23.3, 23.6,24.6, 24.9, 25.3, 25.9, 26.9, 27.3, 27.4, 28.1, and 28.2 degrees 2θ. Inanother embodiment, the anhydrous hydrochloric acid salt of the compoundof Formula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising at least three diffraction angles, when measuredusing Cu K_(α) radiation, selected from a group consisting of about 8.3,8.4, 10.7, 11.3, 15.5, 16.0, 20.0, 20.4, 20.8, 22.6, 23.2, 23.3, 23.6,24.6, 24.9, 25.3, 25.9, 26.9, 27.3, 27.4, 28.1, and 28.2 degrees 2θ.

In still another embodiment, the anhydrous hydrochloric acid salt of thecompound of Formula (I) is characterized by an X-ray powder diffraction(XRPD) pattern comprising diffraction angles, when measured using CuK_(α) radiation, of about 16.0, 20.0, 22.6, 23.3, and 26.9 degrees 2θ.In yet another embodiment, the anhydrous hydrochloric acid salt of thecompound of Formula (I) is characterized by an X-ray powder diffraction(XRPD) pattern substantially in accordance with FIG. 9.

In other embodiments, the anhydrous hydrochloric acid salt of thecompound of Formula (I) is characterized by a Raman spectrum comprisingat least nine peaks at positions selected from a group consisting ofpeaks at about 418, 454, 575, 636, 699, 771, 782, 805, 864, 894, 941,974, 998, 1058, 1116, 1190, 1246, 1273, 1299, 1329, 1356, 1407, 1433,1462, 1489, 1511, 1546, 1562, 1614, 1626, 1667, 1695, 2922, 2950, 2986,3036, 3075, and 3095 cm⁻¹. In another embodiment, the anhydroushydrochloric acid salt of the compound of Formula (I) is characterizedby a Raman spectrum comprising at least eight peaks or at least sevenpeaks or at least six peaks or at least five peaks or at least fourthree peaks at positions selected from a group consisting of peaks atabout 418, 454, 575, 636, 699, 771, 782, 805, 864, 894, 941, 974, 998,1058, 1116, 1190, 1246, 1273, 1299, 1329, 1356, 1407, 1433, 1462, 1489,1511, 1546, 1562, 1614, 1626, 1667, 1695, 2922, 2950, 2986, 3036, 3075,and 3095 cm⁻¹. In another embodiment, the anhydrous hydrochloric acidsalt of the compound of Formula (I) is characterized by a Raman spectrumcomprising at least three peaks at positions selected from a groupconsisting of peaks at about 418, 454, 575, 636, 699, 771, 782, 805,864, 894, 941, 974, 998, 1058, 1116, 1190, 1246, 1273, 1299, 1329, 1356,1407, 1433, 1462, 1489, 1511, 1546, 1562, 1614, 1626, 1667, 1695, 2922,2950, 2986, 3036, 3075, and 3095 cm⁻¹.

In still another embodiment, the anhydrous hydrochloric acid salt of thecompound of Formula (I) is characterized by a Raman spectrum comprisingpeaks at about 771, 805, 998, 1058, 1246, 1329, 1614, 1626, 2922, 2950,and 3036 cm⁻¹. In yet another embodiment, the anhydrous hydrochloricacid salt of the compound of Formula (I) is characterized by a Ramanspectrum substantially in accordance with FIG. 10.

In further embodiments, the anhydrous hydrochloric acid salt of thecompound of Formula (I) is characterized by a differential scanningcalorimetry trace substantially in accordance with FIG. 11 and/or athermogravimetric analysis trace substantially in accordance with FIG.12.

In still further embodiments, as a person having ordinary skill in theart will understand, the anhydrous hydrochloric acid salt of thecompound of Formula (I) is characterized by any combination of theanalytical data characterizing the aforementioned embodiments. Forexample, in one embodiment, the anhydrous hydrochloric acid salt of thecompound of Formula (I) is characterized by an X-ray powder diffraction(XRPD) pattern substantially in accordance with FIG. 9 and a Ramanspectrum substantially in accordance with FIG. 10 and a differentialscanning calorimetry trace substantially in accordance with FIG. 11 anda thermogravimetric analysis trace substantially in accordance with FIG.12. In another embodiment, the anhydrous hydrochloric acid salt of thecompound of Formula (I) is characterized by an X-ray powder diffraction(XRPD) pattern substantially in accordance with FIG. 9 and a Ramanspectrum substantially in accordance with FIG. 10. In anotherembodiment, the anhydrous hydrochloric acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern substantially in accordance with FIG. 9 and a differentialscanning calorimetry trace substantially in accordance with FIG. 11. Inanother embodiment, the anhydrous hydrochloric acid salt of the compoundof Formula (I) is characterized by an X-ray powder diffraction (XRPD)pattern substantially in accordance with FIG. 9 and a thermogravimetricanalysis trace substantially in accordance with FIG. 12. In anotherembodiment, the anhydrous hydrochloric acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising diffraction angles, when measured using Cu K_(α)radiation, of about 16.0, 20.0, 22.6, 23.3, and 26.9 degrees 2θ, and aRaman spectrum comprising peaks at about 771, 805, 998, 1058, 1246,1329, 1614, 1626, 2922, 2950, and 3036 cm⁻¹. In another embodiment, theanhydrous hydrochloric acid salt of the compound of Formula (I) ischaracterized by an X-ray powder diffraction (XRPD) pattern comprisingdiffraction angles, when measured using Cu K_(α) radiation, of about16.0, 20.0, 22.6, 23.3, and 26.9 degrees 2θ, and a differential scanningcalorimetry trace substantially in accordance with FIG. 11. In anotherembodiment, the anhydrous hydrochloric acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising diffraction angles, when measured using Cu K_(α)radiation, of about 16.0, 20.0, 22.6, 23.3, and 26.9 degrees 2θ, and athermogravimetric analysis trace substantially in accordance with FIG.12.

In some embodiments, a crystalline form ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate (the aspartic acid salt of the compound of Formula (I)) ischaracterized by an X-ray powder diffraction (XRPD) pattern comprisingat least nine diffraction angles, when measured using Cu K_(α)radiation, selected from a group consisting of about 6.7, 7.0, 7.6,11.8, 13.9, 14.8, 15.6, 15.8, 16.2, 17.7, 18.4, 18.7, 19.1, 19.2, 20.1,20.6, 21.0, 21.1, 21.2, 21.7, 22.1, 22.8, 23.0, 23.1, 23.3, 23.7, 23.8,25.0, 25.1, 25.4, 25.5, 25.7, 26.2, 27.4, 28.2, 31.2, 35.9, and 36.0degrees 2θ. In another embodiment, the aspartic acid salt of thecompound of Formula (I) is characterized by an X-ray powder diffraction(XRPD) pattern comprising at least eight diffraction angles or at leastseven diffraction angles or at least six diffraction angles or at leastfive diffraction angles or at least four diffraction angles, whenmeasured using Cu K_(α) radiation, selected from a group consisting ofabout 6.7, 7.0, 7.6, 11.8, 13.9, 14.8, 15.6, 15.8, 16.2, 17.7, 18.4,18.7, 19.1, 19.2, 20.1, 20.6, 21.0, 21.1, 21.2, 21.7, 22.1, 22.8, 23.0,23.1, 23.3, 23.7, 23.8, 25.0, 25.1, 25.4, 25.5, 25.7, 26.2, 27.4, 28.2,31.2, 35.9, and 36.0 degrees 2θ. In another embodiment, the asparticacid salt of the compound of Formula (I) is characterized by an X-raypowder diffraction (XRPD) pattern comprising at least three diffractionangles, when measured using Cu K_(α) radiation, selected from a groupconsisting of about 6.7, 7.0, 7.6, 11.8, 13.9, 14.8, 15.6, 15.8, 16.2,17.7, 18.4, 18.7, 19.1, 19.2, 20.1, 20.6, 21.0, 21.1, 21.2, 21.7, 22.1,22.8, 23.0, 23.1, 23.3, 23.7, 23.8, 25.0, 25.1, 25.4, 25.5, 25.7, 26.2,27.4, 28.2, 31.2, 35.9, and 36.0 degrees 2θ.

In still another embodiment, the aspartic acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising diffraction angles, when measured using Cu K_(α)radiation, of about 7.0, 7.6, 11.8, 16.2, 20.6, 21.7, and 23.8 degrees2θ. In yet another embodiment, the aspartic acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern substantially in accordance with FIG. 13.

In other embodiments, the aspartic acid salt of the compound of Formula(I) is characterized by a Raman spectrum comprising at least nine peaksat positions selected from a group consisting of peaks at about 452,574, 691, 770, 807, 1000, 1037, 1106, 1162, 1237, 1274, 1332, 1364,1471, 1487, 1530, 1627, 1705, 2918, and 3073 cm⁻¹. In anotherembodiment, the aspartic acid salt of the compound of Formula (I) ischaracterized by a Raman spectrum comprising at least eight peaks or atleast seven peaks or at least six peaks or at least five peaks or atleast four three peaks at positions selected from a group consisting ofpeaks at about 452, 574, 691, 770, 807, 1000, 1037, 1106, 1162, 1237,1274, 1332, 1364, 1471, 1487, 1530, 1627, 1705, 2918, and 3073 cm⁻¹. Inanother embodiment, the aspartic acid salt of the compound of Formula(I) is characterized by a Raman spectrum comprising at least three peaksat positions selected from a group consisting of peaks at about 452,574, 691, 770, 807, 1000, 1037, 1106, 1162, 1237, 1274, 1332, 1364,1471, 1487, 1530, 1627, 1705, 2918, and 3073 cm⁻¹.

In still another embodiment, the aspartic acid salt of the compound ofFormula (I) is characterized by a Raman spectrum comprising peaks atabout 770, 807, 1000, 1237, 1274, 1332, 1364, 1471, 1627, 2918, and 3073cm⁻¹. In yet another embodiment, the aspartic acid salt of the compoundof Formula (I) is characterized by a Raman spectrum substantially inaccordance with FIG. 14.

In further embodiments, the aspartic acid salt of the compound ofFormula (I) is characterized by a differential scanning calorimetrytrace substantially in accordance with FIG. 15 and/or athermogravimetric analysis trace substantially in accordance with FIG.16.

In still further embodiments, as a person having ordinary skill in theart will understand, the aspartic acid salt of the compound of Formula(I) is characterized by any combination of the analytical datacharacterizing the aforementioned embodiments. For example, in oneembodiment, the aspartic acid salt of the compound of Formula (I) ischaracterized by an X-ray powder diffraction (XRPD) patternsubstantially in accordance with FIG. 13 and a Raman spectrumsubstantially in accordance with FIG. 14 and a differential scanningcalorimetry trace substantially in accordance with FIG. 15 and athermogravimetric analysis trace substantially in accordance with FIG.16. In another embodiment, the aspartic acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern substantially in accordance with FIG. 13 and a Raman spectrumsubstantially in accordance with FIG. 14. In another embodiment, theaspartic acid salt of the compound of Formula (I) is characterized by anX-ray powder diffraction (XRPD) pattern substantially in accordance withFIG. 13 and a differential scanning calorimetry trace substantially inaccordance with FIG. 15. In another embodiment, the aspartic acid saltof the compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern substantially in accordance with FIG. 13 anda thermogravimetric analysis trace substantially in accordance with FIG.16. In another embodiment, the aspartic acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising diffraction angles, when measured using Cu K_(α)radiation, of about 7.0, 7.6, 11.8, 16.2, 20.6, 21.7, and 23.8 degrees2θ, and a Raman spectrum comprising peaks at about 770, 807, 1000, 1237,1274, 1332, 1364, 1471, 1627, 2918, and 3073 cm⁻¹. In anotherembodiment, the aspartic acid salt of the compound of Formula (I) ischaracterized by an X-ray powder diffraction (XRPD) pattern comprisingdiffraction angles, when measured using Cu K_(α) radiation, of about7.0, 7.6, 11.8, 16.2, 20.6, 21.7, and 23.8 degrees 2θ, and adifferential scanning calorimetry trace substantially in accordance withFIG. 15. In another embodiment, the aspartic acid salt of the compoundof Formula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising diffraction angles, when measured using Cu K_(α)radiation, of about 7.0, 7.6, 11.8, 16.2, 20.6, 21.7, and 23.8 degrees2θ, and a thermogravimetric analysis trace substantially in accordancewith FIG. 16.

In some embodiments, a crystalline form ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate (the hippuric acid salt of the compound of Formula (I)) ischaracterized by an X-ray powder diffraction (XRPD) pattern comprisingat least nine diffraction angles, when measured using Cu K_(α)radiation, selected from a group consisting of about 6.1, 9.2, 10.1,12.0, 12.3, 12.6, 13.8, 14.0, 17.2, 17.3, 18.1, 18.4, 18.9, 19.0, 19.1,19.6, 20.6, 21.1, 21.2, 21.5, 21.7, 22.4, 22.9, 23.5, 23.6, 24.9, 27.2,27.4, 27.6, and 28.0 degrees 2θ. In another embodiment, the hippuricacid salt of the compound of Formula (I) is characterized by an X-raypowder diffraction (XRPD) pattern comprising at least eight diffractionangles or at least seven diffraction angles or at least six diffractionangles or at least five diffraction angles or at least four diffractionangles, when measured using Cu K_(α) radiation, selected from a groupconsisting of about 6.1, 9.2, 10.1, 12.0, 12.3, 12.6, 13.8, 14.0, 17.2,17.3, 18.1, 18.4, 18.9, 19.0, 19.1, 19.6, 20.6, 21.1, 21.2, 21.5, 21.7,22.4, 22.9, 23.5, 23.6, 24.9, 27.2, 27.4, 27.6, and 28.0 degrees 2θ. Inanother embodiment, the hippuric acid salt of the compound of Formula(I) is characterized by an X-ray powder diffraction (XRPD) patterncomprising at least three diffraction angles, when measured using CuK_(α) radiation, selected from a group consisting of about 6.1, 9.2,10.1, 12.0, 12.3, 12.6, 13.8, 14.0, 17.2, 17.3, 18.1, 18.4, 18.9, 19.0,19.1, 19.6, 20.6, 21.1, 21.2, 21.5, 21.7, 22.4, 22.9, 23.5, 23.6, 24.9,27.2, 27.4, 27.6, and 28.0 degrees 2θ.

In still another embodiment, the hippuric acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising diffraction angles, when measured using Cu K_(α)radiation, of about 6.1, 9.2, 12.6, 18.4, 20.6, and 22.4 degrees 2θ. Inyet another embodiment, the hippuric acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern substantially in accordance with FIG. 17.

In other embodiments, the hippuric acid salt of the compound of Formula(I) is characterized by a Raman spectrum comprising at least nine peaksat positions selected from a group consisting of peaks at about 557,618, 694, 771, 809, 917, 997, 1042, 1108, 1236, 1272, 1335, 1366, 1467,1537, 1575, 1601, 1630, 1695, 2944, and 3071 cm⁻¹. In anotherembodiment, the hippuric acid salt of the compound of Formula (I) ischaracterized by a Raman spectrum comprising at least eight peaks or atleast seven peaks or at least six peaks or at least five peaks or atleast four three peaks at positions selected from a group consisting ofpeaks at about 557, 618, 694, 771, 809, 917, 997, 1042, 1108, 1236,1272, 1335, 1366, 1467, 1537, 1575, 1601, 1630, 1695, 2944, and 3071cm⁻¹. In another embodiment, the hippuric acid salt of the compound ofFormula (I) is characterized by a Raman spectrum comprising at leastthree peaks at positions selected from a group consisting of peaks atabout 557, 618, 694, 771, 809, 917, 997, 1042, 1108, 1236, 1272, 1335,1366, 1467, 1537, 1575, 1601, 1630, 1695, 2944, and 3071 cm⁻¹.

In still another embodiment, the hippuric acid salt of the compound ofFormula (I) is characterized by a Raman spectrum comprising peaks atabout 809, 997, 1236, 1272, 1335, 1366, 1601, 1630, 2944, and 3071 cm⁻¹.In yet another embodiment, the hippuric acid salt of the compound ofFormula (I) is characterized by a Raman spectrum substantially inaccordance with FIG. 18.

In further embodiments, the hippuric acid salt of the compound ofFormula (I) is characterized by a differential scanning calorimetrytrace substantially in accordance with FIG. 19 and/or athermogravimetric analysis trace substantially in accordance with FIG.20.

In still further embodiments, as a person having ordinary skill in theart will understand, the hippuric acid salt of the compound of Formula(I) is characterized by any combination of the analytical datacharacterizing the aforementioned embodiments. For example, in oneembodiment, the hippuric acid salt of the compound of Formula (I) ischaracterized by an X-ray powder diffraction (XRPD) patternsubstantially in accordance with FIG. 17 and a Raman spectrumsubstantially in accordance with FIG. 18 and a differential scanningcalorimetry trace substantially in accordance with FIG. 19 and athermogravimetric analysis trace substantially in accordance with FIG.20. In another embodiment, the hippuric acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern substantially in accordance with FIG. 17 and a Raman spectrumsubstantially in accordance with FIG. 18. In another embodiment, thehippuric acid salt of the compound of Formula (I) is characterized by anX-ray powder diffraction (XRPD) pattern substantially in accordance withFIG. 17 and a differential scanning calorimetry trace substantially inaccordance with FIG. 19. In another embodiment, the hippuric acid saltof the compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern substantially in accordance with FIG. 17 anda thermogravimetric analysis trace substantially in accordance with FIG.20. In another embodiment, the hippuric acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising diffraction angles, when measured using Cu K_(α)radiation, of about 6.1, 9.2, 12.6, 18.4, 20.6, and 22.4 degrees 2θ, anda Raman spectrum comprising peaks at about 809, 997, 1236, 1272, 1335,1366, 1601, 1630, 2944, and 3071 cm⁻¹. In another embodiment, thehippuric acid salt of the compound of Formula (I) is characterized by anX-ray powder diffraction (XRPD) pattern comprising diffraction angles,when measured using Cu K_(α) radiation, of about 6.1, 9.2, 12.6, 18.4,20.6, and 22.4 degrees 2θ, and a differential scanning calorimetry tracesubstantially in accordance with FIG. 19. In another embodiment, thehippuric acid salt of the compound of Formula (I) is characterized by anX-ray powder diffraction (XRPD) pattern comprising diffraction angles,when measured using Cu K_(α) radiation, of about 6.1, 9.2, 12.6, 18.4,20.6, and 22.4 degrees 2θ, and a thermogravimetric analysis tracesubstantially in accordance with FIG. 20.

In some embodiments, a crystalline form ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate (the phosphoric acid salt of the compound of Formula (I)) ischaracterized by an X-ray powder diffraction (XRPD) pattern comprisingat least nine diffraction angles, when measured using Cu K_(α)radiation, selected from a group consisting of about 5.5, 5.6, 7.7, 9.9,11.2, 15.4, 16.0, 16.8, 18.2, 19.9, 20.3, 23.9, 24.2, 24.4, 26.5, 26.7,27.0, and 28.7 degrees 2θ. In another embodiment, the phosphoric acidsalt of the compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern comprising at least eight diffraction anglesor at least seven diffraction angles or at least six diffraction anglesor at least five diffraction angles or at least four diffraction angles,when measured using Cu K_(α) radiation, selected from a group consistingof about 5.5, 5.6, 7.7, 9.9, 11.2, 15.4, 16.0, 16.8, 18.2, 19.9, 20.3,23.9, 24.2, 24.4, 26.5, 26.7, 27.0, and 28.7 degrees 2θ. In anotherembodiment, the phosphoric acid salt of the compound of Formula (I) ischaracterized by an X-ray powder diffraction (XRPD) pattern comprisingat least three diffraction angles, when measured using Cu K_(α)radiation, selected from a group consisting of about 5.5, 5.6, 7.7, 9.9,11.2, 15.4, 16.0, 16.8, 18.2, 19.9, 20.3, 23.9, 24.2, 24.4, 26.5, 26.7,27.0, and 28.7 degrees 2θ.

In still another embodiment, the phosphoric acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising diffraction angles, when measured using Cu K_(α)radiation, of about 9.9, 16.8, 19.9, 20.3, 24.2, 26.5, and 27.0 degrees2θ. In yet another embodiment, the phosphoric acid salt of the compoundof Formula (I) is characterized by an X-ray powder diffraction (XRPD)pattern substantially in accordance with FIG. 21.

In other embodiments, the phosphoric acid salt of the compound ofFormula (I) is characterized by a Raman spectrum comprising at leastnine peaks at positions selected from a group consisting of peaks atabout 461, 485, 529, 577, 638, 696, 732, 773, 786, 806, 867, 889, 1002,1036, 1187, 1243, 1276, 1296, 1326, 1358, 1375, 1442, 1466, 1510, 1532,1580, 1625, 1698, 2936, 2964, and 3069 cm⁻¹. In another embodiment, thephosphoric acid salt of the compound of Formula (I) is characterized bya Raman spectrum comprising at least eight peaks or at least seven peaksor at least six peaks or at least five peaks or at least four threepeaks at positions selected from a group consisting of peaks at about461, 485, 529, 577, 638, 696, 732, 773, 786, 806, 867, 889, 1002, 1036,1187, 1243, 1276, 1296, 1326, 1358, 1375, 1442, 1466, 1510, 1532, 1580,1625, 1698, 2936, 2964, and 3069 cm⁻¹. In another embodiment, thephosphoric acid salt of the compound of Formula (I) is characterized bya Raman spectrum comprising at least three peaks at positions selectedfrom a group consisting of peaks at about 461, 485, 529, 577, 638, 696,732, 773, 786, 806, 867, 889, 1002, 1036, 1187, 1243, 1276, 1296, 1326,1358, 1375, 1442, 1466, 1510, 1532, 1580, 1625, 1698, 2936, 2964, and3069 cm⁻¹.

In still another embodiment, the phosphoric acid salt of the compound ofFormula (I) is characterized by a Raman spectrum comprising peaks atabout 786, 806, 1002, 1036, 1243, 1296, 1326, 1375, 1625, 2936, and 2964cm⁻¹. In yet another embodiment, the phosphoric acid salt of thecompound of Formula (I) is characterized by a Raman spectrumsubstantially in accordance with FIG. 22.

In further embodiments, the phosphoric acid salt of the compound ofFormula (I) is characterized by a differential scanning calorimetrytrace substantially in accordance with FIG. 23 and/or athermogravimetric analysis trace substantially in accordance with FIG.24.

In still further embodiments, as a person having ordinary skill in theart will understand, the phosphoric acid salt of the compound of Formula(I) is characterized by any combination of the analytical datacharacterizing the aforementioned embodiments. For example, in oneembodiment, the phosphoric acid salt of the compound of Formula (I) ischaracterized by an X-ray powder diffraction (XRPD) patternsubstantially in accordance with FIG. 21 and a Raman spectrumsubstantially in accordance with FIG. 22 and a differential scanningcalorimetry trace substantially in accordance with FIG. 23 and athermogravimetric analysis trace substantially in accordance with FIG.24. In another embodiment, the phosphoric acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern substantially in accordance with FIG. 21 and a Raman spectrumsubstantially in accordance with FIG. 22. In another embodiment, thephosphoric acid salt of the compound of Formula (I) is characterized byan X-ray powder diffraction (XRPD) pattern substantially in accordancewith FIG. 21 and a differential scanning calorimetry trace substantiallyin accordance with FIG. 23. In another embodiment, the phosphoric acidsalt of the compound of Formula (I) is characterized by an X-ray powderdiffraction (XRPD) pattern substantially in accordance with FIG. 21 anda thermogravimetric analysis trace substantially in accordance with FIG.24. In another embodiment, the phosphoric acid salt of the compound ofFormula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising diffraction angles, when measured using Cu K_(α)radiation, of about 9.9, 16.8, 19.9, 20.3, 24.2, 26.5, and 27.0 degrees2θ, and a Raman spectrum comprising peaks at about 786, 806, 1002, 1036,1243, 1296, 1326, 1375, 1625, 2936, and 2964 cm⁻¹. In anotherembodiment, the phosphoric acid salt of the compound of Formula (I) ischaracterized by an X-ray powder diffraction (XRPD) pattern comprisingdiffraction angles, when measured using Cu K_(α) radiation, of about9.9, 16.8, 19.9, 20.3, 24.2, 26.5, and 27.0 degrees 2θ, and adifferential scanning calorimetry trace substantially in accordance withFIG. 23. In another embodiment, the phosphoric acid salt of the compoundof Formula (I) is characterized by an X-ray powder diffraction (XRPD)pattern comprising diffraction angles, when measured using Cu K_(α)radiation, of about 9.9, 16.8, 19.9, 20.3, 24.2, 26.5, and 27.0 degrees2θ, and a thermogravimetric analysis trace substantially in accordancewith FIG. 24.

An XRPD pattern will be understood to comprise a diffraction angle(expressed in degrees 2θ) of “about” a value specified herein when theXRPD pattern comprises a diffraction angle within ±0.3 degrees 2θ of thespecified value. Further, it is well known and understood to thoseskilled in the art that the apparatus employed, humidity, temperature,orientation of the powder crystals, and other parameters involved inobtaining an X-ray powder diffraction (XRPD) pattern may cause somevariability in the appearance, intensities, and positions of the linesin the diffraction pattern. An X-ray powder diffraction pattern that is“substantially in accordance” with that of FIG. 1, 5, 9, 13, 17, or 21provided herein is an XRPD pattern that would be considered by oneskilled in the art to represent a compound possessing the same crystalform as the compound that provided the XRPD pattern of FIG. 1, 5, 9, 13,17, or 21. That is, the XRPD pattern may be identical to that of FIG. 1,5, 9, 13, 17, or 21, or more likely it may be somewhat different. Suchan XRPD pattern may not necessarily show each of the lines of any one ofthe diffraction patterns presented herein, and/or may show a slightchange in appearance, intensity, or a shift in position of said linesresulting from differences in the conditions involved in obtaining thedata. A person skilled in the art is capable of determining if a sampleof a crystalline compound has the same form as, or a different formfrom, a form disclosed herein by comparison of their XRPD patterns. Forexample, one skilled in the art can overlay an XRPD pattern of a sampleofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride, with FIG. 1 and, using expertise and knowledge in theart, readily determine whether the XRPD pattern of the sample issubstantially in accordance with the XRPD pattern of hydrate 1 of thehydrochloric acid salt of the compound of Formula (I) disclosed herein.If the XRPD pattern is substantially in accordance with FIG. 1, thesample form can be readily and accurately identified as having the sameform as hydrate 1 of the hydrochloric acid salt of the compound ofFormula (I) disclosed herein. Similarly, if an XRPD pattern of a sampleofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is substantially in accordance with FIG. 5, the sampleform can be readily and accurately identified as having the same form ashydrate 2 of the hydrochloric acid salt of the compound of Formula (I)disclosed herein. Similarly, if an XRPD pattern of a sample ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is substantially in accordance with FIG. 9, the sampleform can be readily and accurately identified as having the same form asthe anhydrous hydrochloric acid salt of the compound of Formula (I)disclosed herein. Similarly, if an XRPD pattern of a sample ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate is substantially in accordance with FIG. 13, the sample formcan be readily and accurately identified as having the same form as theaspartic acid salt of the compound of Formula (I) disclosed herein.Similarly, if an XRPD pattern of a sample ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate is substantially in accordance with FIG. 17, the sample formcan be readily and accurately identified as having the same form as thehippuric acid salt of the compound of Formula (I) disclosed herein.Similarly, if an XRPD pattern of a sample ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate is substantially in accordance with FIG. 21, the sample formcan be readily and accurately identified as having the same form as thephosphoric acid salt of the compound of Formula (I) disclosed herein.

A Raman spectrum will be understood to comprise a peak (expressed incm⁻¹) of “about” a value specified herein when the Raman spectrumcomprises a peak within ±5.0 cm⁻¹ of the specified value. Further, it isalso well known and understood to those skilled in the art that theapparatus employed, humidity, temperature, orientation of the powdercrystals, and other parameters involved in obtaining a Raman spectrummay cause some variability in the appearance, intensities, and positionsof the peaks in the spectrum. A Raman spectrum that is “substantially inaccordance” with that of FIG. 2, 6, 10, 14, 18, or 22 provided herein isa Raman spectrum that would be considered by one skilled in the art torepresent a compound possessing the same crystal form as the compoundthat provided the Raman spectrum of FIG. 2, 6, 10, 14, 18, or 22. Thatis, the Raman spectrum may be identical to that of FIG. 2, 6, 10, 14,18, or 22, or more likely it may be somewhat different. Such a Ramanspectrum may not necessarily show each of the peaks of any one of thespectra presented herein, and/or may show a slight change in appearance,intensity, or a shift in position of said peaks resulting fromdifferences in the conditions involved in obtaining the data. A personskilled in the art is capable of determining if a sample of acrystalline compound has the same form as, or a different form from, aform disclosed herein by comparison of their Raman spectra. For example,one skilled in the art can overlay a Raman spectrum of a sample ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride, with FIG. 2 and, using expertise and knowledge in theart, readily determine whether the Raman spectrum of the sample issubstantially in accordance with the Raman spectrum of hydrate 1 of thehydrochloric acid salt of the compound of Formula (I) disclosed herein.If the Raman spectrum is substantially in accordance with FIG. 6, thesample form can be readily and accurately identified as having the sameform as hydrate 2 of the hydrochloric acid salt of the compound ofFormula (I) disclosed herein. Similarly, if the Raman spectrum issubstantially in accordance with FIG. 10, the sample form can be readilyand accurately identified as having the same form as the anhydroushydrochloric acid salt of the compound of Formula (I) disclosed herein.Similarly, if the Raman spectrum is substantially in accordance withFIG. 14, the sample form can be readily and accurately identified ashaving the same form as the aspartic acid salt of the compound ofFormula (I) disclosed herein. Similarly, if the Raman spectrum issubstantially in accordance with FIG. 18, the sample form can be readilyand accurately identified as having the same form as the hippuric acidsalt of the compound of Formula (I) disclosed herein. Similarly, if theRaman spectrum is substantially in accordance with FIG. 22, the sampleform can be readily and accurately identified as having the same form asthe phosphoric acid salt of the compound of Formula (I) disclosedherein.

It is further understood that a compound or salt of Formula (I) or (II)may exist in tautomeric forms other than that shown in the formula andthese are also included within the scope of the present invention. Forexample, while the compounds of Formula (I) and (II) are depicted ascontaining a pyridin-2-one moiety, the corresponding 2-hydroxypyridinetautomer is also included within the scope of the present invention.

It will be appreciated by those skilled in the art that certainprotected derivatives of compounds of Formula (I) or (II), which may bemade prior to or following a final deprotection stage, may not possesspharmacological activity as such, but may, in certain instances, beadministered orally or parenterally and thereafter metabolized in thebody to form compounds of the invention which are pharmacologicallyactive. Such derivatives may therefore be described as “prodrugs”. Allprotected derivatives and prodrugs of compounds of the invention areincluded within the scope of the invention.

Examples of suitable pro-drugs for the compounds of the presentinvention are described in Drugs of Today, Volume 19, Number 9, 1983, pp499-538 and in Topics in Chemistry, Chapter 31, pp 306-316 and in“Design of Prodrugs” by H. Bundgaard, Elsevier, 1985, Chapter 1. It willfurther be appreciated by those skilled in the art, that certainmoieties, known to those skilled in the art as “pro-moieties”, forexample as described by H. Bundgaard in “Design of Prodrugs” may beplaced on appropriate functionalities when such functionalities arepresent within compounds of the invention. Preferred “pro-moieties” forcompounds of the invention include: ester, carbonate ester, hemi-ester,phosphate ester, nitro ester, sulfate ester, sulfoxide, amide,carbamate, azo-, phosphamide, glycoside, ether, acetal, and ketalderivatives of the compounds of Formula (I) or (II).

Administration of a compound of the invention as a prodrug may enablethe skilled artisan to do one or more of the following: (a) modify theonset of the compound in vivo; (b) modify the duration of action of thecompound in vivo; (c) modify the transportation or distribution of thecompound in vivo; (d) modify the solubility of the compound in vivo; and(e) overcome or overcome a side effect or other difficulty encounteredwith the compound.

The subject invention also includes isotopically-labelled compounds,which are identical to those recited in Formula (I) or (II), but for thefact that one or more atoms are replaced by an atom having an atomicmass or mass number different from the atomic mass or mass numberusually found in nature. Examples of isotopes that can be incorporatedinto compounds of the invention and pharmaceutically acceptable saltsthereof include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorous, sulphur, fluorine, iodine, and chlorine, such as ²H, ³H,¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁷O, ¹⁸O, ³¹P, ³²P, ³⁵S, ¹⁸F, ³⁶Cl, ¹²³I, and ¹²⁵I.

Compounds of the present invention and pharmaceutically acceptable saltsof said compounds that contain the aforementioned isotopes and/or otherisotopes of other atoms are within the scope of the present invention.Isotopically-labelled compounds of the present invention, for examplethose into which radioactive isotopes such as ³H or ¹⁴C areincorporated, are useful in drug and/or substrate tissue distributionassays. Tritiated, i.e., ³H, and carbon-14, i.e., ¹⁴C, isotopes areparticularly preferred for their ease of preparation and detectability.¹¹C and ¹⁸F isotopes are particularly useful in PET (positron emissiontomography), and ¹²⁵I isotopes are particularly useful in SPECT (singlephoton emission computerized tomography), all useful in brain imaging.Further, substitution with heavier isotopes such as deuterium, i.e., ²H,can afford certain therapeutic advantages resulting from greatermetabolic stability, for example increased in vivo half-life or reduceddosage requirements and, hence, may be preferred in some circumstances.Isotopically labelled compounds of Formula (I) and following of thisinvention can generally be prepared by carrying out the proceduresdisclosed in the Schemes and/or in the Examples below, by substituting areadily available isotopically labelled reagent for a non-isotopicallylabelled reagent.

Definitions

Terms are used within their accepted meanings. The following definitionsare meant to clarify, but not limit, the terms defined.

“Pharmaceutically acceptable” refers to those compounds, materials,compositions, and dosage forms which are, within the scope of soundmedical judgment, suitable for use in contact with the tissues of humanbeings and animals without excessive toxicity, irritation, or otherproblem or complication, commensurate with a reasonable benefit/riskratio.

As used herein, the term “pharmaceutically acceptable salts” refers tosalts that retain the desired biological activity of the subjectcompound and exhibit minimal undesired toxicological effects. Thesepharmaceutically acceptable salts may be prepared in situ during thefinal isolation and purification of the compound, or by separatelyreacting the purified compound in its free acid or free base form with asuitable base or acid, respectively.

Pharmaceutical Compositions

The invention further provides a pharmaceutical composition (alsoreferred to as pharmaceutical formulation) comprising a compound ofFormula (I) or (II) or a pharmaceutically acceptable salt thereof, andone or more excipients (also referred to as carriers and/or diluents inthe pharmaceutical arts). The excipients are pharmaceutically acceptablein the sense of being compatible with the other ingredients of theformulation and not deleterious to the recipient thereof (i.e., thepatient).

Suitable pharmaceutically acceptable excipients include the followingtypes of excipients: diluents, fillers, binders, disintegrants,lubricants, glidants, granulating agents, coating agents, wettingagents, solvents, co-solvents, suspending agents, emulsifiers,sweeteners, flavoring agents, flavor masking agents, coloring agents,anticaking agents, hemectants, chelating agents, plasticizers, viscosityincreasing agents, antioxidants, preservatives, stabilizers,surfactants, and buffering agents. The skilled artisan will appreciatethat certain pharmaceutically acceptable excipients may serve more thanone function and may serve alternative functions depending on how muchof the excipient is present in the formulation and what otheringredients are present in the formulation.

Skilled artisans possess the knowledge and skill in the art to enablethem to select suitable pharmaceutically acceptable excipients inappropriate amounts for use in the invention. In addition, there are anumber of resources that are available to the skilled artisan whichdescribe pharmaceutically acceptable excipients and may be useful inselecting suitable pharmaceutically acceptable excipients. Examplesinclude Remington's Pharmaceutical Sciences (Mack Publishing Company),The Handbook of Pharmaceutical Additives (Gower Publishing Limited), andThe Handbook of Pharmaceutical Excipients (the American PharmaceuticalAssociation and the Pharmaceutical Press).

The pharmaceutical compositions of the invention are prepared usingtechniques and methods known to those skilled in the art. Some of themethods commonly used in the art are described in Remington'sPharmaceutical Sciences (Mack Publishing Company).

In accordance with another aspect of the invention there is provided aprocess for the preparation of a pharmaceutical composition comprisingmixing (or admixing) a compound of Formula (I) or (II) or apharmaceutically acceptable salt thereof, with at least one excipient.

Pharmaceutical compositions may be in unit dose form containing apredetermined amount of active ingredient per unit dose. Such a unit maycontain a therapeutically effective dose of the compound of Formula (I)or (II) or a pharmaceutically acceptable salt thereof, or a fraction ofa therapeutically effective dose such that multiple unit dosage formsmight be administered at a given time to achieve the desiredtherapeutically effective dose. Preferred unit dosage formulations arethose containing a daily dose or sub-dose, as herein above recited, oran appropriate fraction thereof, of an active ingredient. Furthermore,such pharmaceutical compositions may be prepared by any of the methodswell-known in the pharmacy art.

Pharmaceutical compositions may be adapted for administration by anyappropriate route, for example, by oral (including buccal orsublingual), rectal, nasal, topical (including buccal, sublingual, ortransdermal), vaginal, or parenteral (including subcutaneous,intramuscular, intravenous, or intradermal) routes. Such compositionsmay be prepared by any method known in the art of pharmacy, for example,by bringing into association the active ingredient with theexcipient(s).

When adapted for oral administration, pharmaceutical compositions may bein discrete units such as tablets or capsules, powders or granules,solutions or suspensions in aqueous or non-aqueous liquids, edible foamsor whips, oil-in-water liquid emulsions or water-in-oil liquidemulsions. The compound or salt thereof of the invention or thepharmaceutical composition of the invention may also be incorporatedinto a candy, a wafer, and/or tongue tape formulation for administrationas a “quick-dissolve” medicine.

For instance, for oral administration in the form of a tablet orcapsule, the active drug component can be combined with an oral,non-toxic pharmaceutically acceptable inert carrier such as ethanol,glycerol, water, and the like. Powders or granules are prepared bycomminuting the compound to a suitable fine size and mixing with asimilarly comminuted pharmaceutical carrier such as an ediblecarbohydrate, as, for example, starch or mannitol Flavoring,preservative, dispersing, and coloring agents can also be present.

Capsules are made by preparing a powder mixture, as described above, andfilling formed gelatin or non-gelatinous sheaths. Glidants andlubricants such as colloidal silica, talc, magnesium stearate, calciumstearate, solid polyethylene glycol can be added to the powder mixturebefore the filling operation. A disintegrating or solubilizing agentsuch as agar-agar, calcium carbonate, or sodium carbonate can also beadded to improve the availability of the medicine when the capsule isingested.

Moreover, when desired or necessary, suitable binders, lubricants,disintegrating agents, and coloring agents can also be incorporated intothe mixture. Suitable binders include starch, gelatin, natural sugars,such as glucose or beta-lactose, corn sweeteners, natural and syntheticgums such as acacia, tragacanth, sodium alginate,carboxymethylcellulose, polyethylene glycol, waxes, and the like.Lubricants used in these dosage forms include sodium oleate, sodiumstearate, magnesium stearate, sodium benzoate, sodium acetate, sodiumchloride, and the like. Disintegrators include, without limitation,starch, methylcellulose, agar, bentonite, xanthan gum, and the like.

Tablets are formulated, for example, by preparing a powder mixture,granulating or slugging, adding a lubricant and disintegrant, andpressing into tablets. A powder mixture is prepared by mixing thecompound, suitably comminuted, with a diluent or base as describedabove, and optionally, with a binder such as carboxymethylcellulose, andaliginate, gelatin, or polyvinyl pyrrolidone, a solution retardant suchas paraffin, a resorption accelerator such as a quaternary salt, and/oran absorption agent such as bentonite, kaolin, or dicalcium phosphate.The powder mixture can be granulated by wetting a binder such as asyrup, starch paste, acadia mucilage, or solutions of cellulosic orpolymeric materials and forcing through a screen. As an alternative togranulating, the powder mixture can be run through a tablet machine,resulting in imperfectly formed slugs broken into granules. The granulescan be lubricated to prevent sticking to the tablet forming dies bymeans of the addition of stearic acid, a stearate salt, talc, or mineraloil. The lubricated mixture is then compressed into tablets. Thecompound or salt of the present invention can also be combined with afree-flowing inert carrier and compressed into tablets directly withoutgoing through the granulating or slugging steps. A clear opaqueprotective coating consisting of a sealing coat of shellac, a coating ofsugar, or polymeric material, and a polish coating of wax can beprovided. Dyestuffs can be added to these coatings to distinguishdifferent dosages.

Oral fluids such as solutions, syrups, and elixirs can be prepared indosage unit form so that a given quantity contains a predeterminedamount of active ingredient. Syrups can be prepared by dissolving thecompound or salt thereof of the invention in a suitably flavouredaqueous solution, while elixirs are prepared through the use of anon-toxic alcoholic vehicle. Suspensions can be formulated by dispersingthe compound or salt of the invention in a non-toxic vehicle.Solubilizers and emulsifiers, such as ethoxylated isostearyl alcoholsand polyoxyethylene sorbitol ethers, preservatives, flavor additivessuch as peppermint oil, natural sweeteners, saccharin, or otherartificial sweeteners, and the like, can also be added.

Where appropriate, dosage unit formulations for oral administration canbe microencapsulated. The formulation can also be prepared to prolong orsustain the release as, for example, by coating or embedding particulatematerial in polymers, wax, or the like.

In the present invention, tablets and capsules are preferred fordelivery of the pharmaceutical composition.

In certain embodiments, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride. In another embodiment, this invention relates to apharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein at least 10% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present as hydrate 1 of the hydrochloric acid salt ofthe compound of Formula (I) described herein. In another embodiment,this invention relates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein at least 20% by weight, or at least 30% by weight,or at least 40% by weight, or at least 50% by weight, or at least 60% byweight, or at least 70% by weight, or at least 80% by weight, or atleast 90% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present as hydrate 1 of the hydrochloric acid salt ofthe compound of Formula (I) described herein. In another embodiment,this invention relates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein at least 95% by weight, or at least 96% by weight,or at least 97% by weight, or at least 98% by weight, or at least 99% byweight, or at least 99.5% by weight, or at least 99.8% by weight, or atleast 99.9% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present as hydrate 1 of the hydrochloric acid salt ofthe compound of Formula (I) described herein.

In another embodiment, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein at least 10% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present as hydrate 2 of the hydrochloric acid salt ofthe compound of Formula (I) described herein. In another embodiment,this invention relates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein at least 20% by weight, or at least 30% by weight,or at least 40% by weight, or at least 50% by weight, or at least 60% byweight, or at least 70% by weight, or at least 80% by weight, or atleast 90% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present as hydrate 2 of the hydrochloric acid salt ofthe compound of Formula (I) described herein. In another embodiment,this invention relates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein at least 95% by weight, or at least 96% by weight,or at least 97% by weight, or at least 98% by weight, or at least 99% byweight, or at least 99.5% by weight, or at least 99.8% by weight, or atleast 99.9% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present as hydrate 2 of the hydrochloric acid salt ofthe compound of Formula (I) described herein.

In another embodiment, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein at least 10% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present as the anhydrous hydrochloric acid salt of thecompound of Formula (I) described herein. In another embodiment, thisinvention relates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein at least 20% by weight, or at least 30% by weight,or at least 40% by weight, or at least 50% by weight, or at least 60% byweight, or at least 70% by weight, or at least 80% by weight, or atleast 90% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present as the anhydrous hydrochloric acid salt of thecompound of Formula (I) described herein. In another embodiment, thisinvention relates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein at least 95% by weight, or at least 96% by weight,or at least 97% by weight, or at least 98% by weight, or at least 99% byweight, or at least 99.5% by weight, or at least 99.8% by weight, or atleast 99.9% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present as the anhydrous hydrochloric acid salt of thecompound of Formula (I) described herein.

In certain embodiments, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate. In another embodiment, this invention relates to apharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate wherein at least 10% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate is present as the aspartic acid salt of the compound ofFormula (I) described herein. In another embodiment, this inventionrelates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate wherein at least 20% by weight, or at least 30% by weight, orat least 40% by weight, or at least 50% by weight, or at least 60% byweight, or at least 70% by weight, or at least 80% by weight, or atleast 90% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate is present as the aspartic acid salt of the compound ofFormula (I) described herein. In another embodiment, this inventionrelates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate wherein at least 95% by weight, or at least 96% by weight, orat least 97% by weight, or at least 98% by weight, or at least 99% byweight, or at least 99.5% by weight, or at least 99.8% by weight, or atleast 99.9% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate is present as the aspartic acid salt of the compound ofFormula (I) described herein.

In certain embodiments, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate. In another embodiment, this invention relates to apharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate wherein at least 10% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate is present as the hippuric acid salt of the compound ofFormula (I) described herein. In another embodiment, this inventionrelates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate wherein at least 20% by weight, or at least 30% by weight, orat least 40% by weight, or at least 50% by weight, or at least 60% byweight, or at least 70% by weight, or at least 80% by weight, or atleast 90% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate is present as the hippuric acid salt of the compound ofFormula (I) described herein. In another embodiment, this inventionrelates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate wherein at least 95% by weight, or at least 96% by weight, orat least 97% by weight, or at least 98% by weight, or at least 99% byweight, or at least 99.5% by weight, or at least 99.8% by weight, or atleast 99.9% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate is present as the hippuric acid salt of the compound ofFormula (I) described herein.

In certain embodiments, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate. In another embodiment, this invention relates to apharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate wherein at least 10% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate is present as the phosphoric acid salt of the compound ofFormula (I) described herein. In another embodiment, this inventionrelates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate wherein at least 20% by weight, or at least 30% by weight, orat least 40% by weight, or at least 50% by weight, or at least 60% byweight, or at least 70% by weight, or at least 80% by weight, or atleast 90% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate is present as the phosphoric acid salt of the compound ofFormula (I) described herein. In another embodiment, this inventionrelates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate wherein at least 95% by weight, or at least 96% by weight, orat least 97% by weight, or at least 98% by weight, or at least 99% byweight, or at least 99.5% by weight, or at least 99.8% by weight, or atleast 99.9% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate is present as the phosphoric acid salt of the compound ofFormula (I) described herein.

In another embodiment, this invention relates to a pharmaceuticalcomposition comprising a pharmaceutically acceptable salt ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide,in particular the hydrochloric acid salt, aspartic acid salt, hippuricacid salt, or phosphoric acid salt, wherein not more than 90% by weightof the salt is amorphous. In another embodiment, this invention relatesto a pharmaceutical composition comprising a pharmaceutically acceptablesalt ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide,in particular the hydrochloric acid salt, aspartic acid salt, hippuricacid salt, or phosphoric acid salt, wherein not more than 80% by weight,or not more than 70% by weight, or not more than 60% by weight, or notmore than 50% by weight, or not more than 40% by weight, or not morethan 30% by weight, or not more than 20% by weight, or not more than 10%by weight of the salt is amorphous. In another embodiment, thisinvention relates to a pharmaceutical composition comprising apharmaceutically acceptable salt ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide,in particular the hydrochloric acid salt, aspartic acid salt, hippuricacid salt, or phosphoric acid salt, wherein not more than 5% by weight,or not more than 4% by weight, or not more than 3% by weight, or notmore than 2% by weight, or not more than 1% by weight, or not more than0.5% by weight, or not more than 0.2% by weight, or not more than 0.1%by weight of the salt is amorphous.

In another embodiment, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein not more than 90% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present in a form other than hydrate 1 of thehydrochloric acid salt of the compound of Formula (I) described herein.In another embodiment, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein not more than 80% by weight, or not more than 70%by weight, or not more than 60% by weight, or not more than 50% byweight, or not more than 40% by weight, or not more than 30% by weight,or not more than 20% by weight, or not more than 10% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present in a form other than hydrate 1 of thehydrochloric acid salt of the compound of Formula (I) described herein.In another embodiment, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein not more than 5% by weight, or not more than 4% byweight, or not more than 3% by weight, or not more than 2% by weight, ornot more than 1% by weight, or not more than 0.5% by weight, or not morethan 0.2% by weight, or not more than 0.1% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present in a form other than hydrate 1 of thehydrochloric acid salt of the compound of Formula (I) described herein.

In another embodiment, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein not more than 90% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present in a form other than hydrate 2 of thehydrochloric acid salt of the compound of Formula (I) described herein.In another embodiment, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein not more than 80% by weight, or not more than 70%by weight, or not more than 60% by weight, or not more than 50% byweight, or not more than 40% by weight, or not more than 30% by weight,or not more than 20% by weight, or not more than 10% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present in a form other than hydrate 2 of thehydrochloric acid salt of the compound of Formula (I) described herein.In another embodiment, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein not more than 5% by weight, or not more than 4% byweight, or not more than 3% by weight, or not more than 2% by weight, ornot more than 1% by weight, or not more than 0.5% by weight, or not morethan 0.2% by weight, or not more than 0.1% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present in a form other than hydrate 2 of thehydrochloric acid salt of the compound of Formula (I) described herein.

In another embodiment, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein not more than 90% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present in a form other than the anhydrous hydrochloricacid salt of the compound of Formula (I) described herein. In anotherembodiment, this invention relates to a pharmaceutical compositioncomprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein not more than 80% by weight, or not more than 70%by weight, or not more than 60% by weight, or not more than 50% byweight, or not more than 40% by weight, or not more than 30% by weight,or not more than 20% by weight, or not more than 10% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present in a form other than the anhydrous hydrochloricacid salt of the compound of Formula (I) described herein. In anotherembodiment, this invention relates to a pharmaceutical compositioncomprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride wherein not more than 5% by weight, or not more than 4% byweight, or not more than 3% by weight, or not more than 2% by weight, ornot more than 1% by weight, or not more than 0.5% by weight, or not morethan 0.2% by weight, or not more than 0.1% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride is present in a form other than the anhydrous hydrochloricacid salt of the compound of Formula (I) described herein.

In another embodiment, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate wherein not more than 90% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate is present in a form other than the aspartic acid salt of thecompound of Formula (I) described herein. In another embodiment, thisinvention relates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate wherein not more than 80% by weight, or not more than 70% byweight, or not more than 60% by weight, or not more than 50% by weight,or not more than 40% by weight, or not more than 30% by weight, or notmore than 20% by weight, or not more than 10% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate is present in a form other than the aspartic acid salt of thecompound of Formula (I) described herein. In another embodiment, thisinvention relates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate wherein not more than 5% by weight, or not more than 4% byweight, or not more than 3% by weight, or not more than 2% by weight, ornot more than 1% by weight, or not more than 0.5% by weight, or not morethan 0.2% by weight, or not more than 0.1% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate is present in a form other than the aspartic acid salt of thecompound of Formula (I) described herein.

In another embodiment, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate wherein not more than 90% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate is present in a form other than the hippuric acid salt of thecompound of Formula (I) described herein. In another embodiment, thisinvention relates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate wherein not more than 80% by weight, or not more than 70% byweight, or not more than 60% by weight, or not more than 50% by weight,or not more than 40% by weight, or not more than 30% by weight, or notmore than 20% by weight, or not more than 10% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate is present in a form other than the hippuric acid salt of thecompound of Formula (I) described herein. In another embodiment, thisinvention relates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate wherein not more than 5% by weight, or not more than 4% byweight, or not more than 3% by weight, or not more than 2% by weight, ornot more than 1% by weight, or not more than 0.5% by weight, or not morethan 0.2% by weight, or not more than 0.1% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate is present in a form other than the hippuric acid salt of thecompound of Formula (I) described herein.

In another embodiment, this invention relates to a pharmaceuticalcomposition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate wherein not more than 90% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate is present in a form other than the phosphoric acid salt ofthe compound of Formula (I) described herein. In another embodiment,this invention relates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate wherein not more than 80% by weight, or not more than 70% byweight, or not more than 60% by weight, or not more than 50% by weight,or not more than 40% by weight, or not more than 30% by weight, or notmore than 20% by weight, or not more than 10% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate is present in a form other than the phosphoric acid salt ofthe compound of Formula (I) described herein. In another embodiment,this invention relates to a pharmaceutical composition comprisingN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate wherein not more than 5% by weight, or not more than 4% byweight, or not more than 3% by weight, or not more than 2% by weight, ornot more than 1% by weight, or not more than 0.5% by weight, or not morethan 0.2% by weight, or not more than 0.1% by weight ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate is present in a form other than the phosphoric acid salt ofthe compound of Formula (I) described herein.

As used herein, the term “treatment” refers to alleviating the specifiedcondition, eliminating or reducing one or more symptoms of thecondition, slowing or eliminating the progression of the condition, andpreventing or delaying the reoccurrence of the condition in a previouslyafflicted or diagnosed patient or subject.

The present invention provides a method of treatment in a mammal,especially a human, suffering from irritable bowel syndrome (IBS)including diarrhea-predominant, constipation-predominant or alternatingstool pattern, functional bloating, functional constipation, functionaldiarrhea, unspecified functional bowel disorder, functional abdominalpain syndrome, chronic idiopathic constipation, functional esophagealdisorders, functional gastroduodenal disorders, functional anorectalpain, inflammatory bowel disease, proliferative diseases such asnon-small cell lung cancer, hepatocellular carcinoma, colorectal cancer,medullary thyroid cancer, follicular thyroid cancer, anaplastic thyroidcancer, papillary thyroid cancer, brain tumors, peritoneal cavitycancer, solid tumors, other lung cancer, head and neck cancer, gliomas,neuroblastomas, Von Hippel-Lindau Syndrome and kidney tumors, breastcancer, fallopian tube cancer, ovarian cancer, transitional cell cancer,prostate cancer, caner of the esophagus and gastroesophageal junction,biliary cancer and adenocarcinoma or a combination thereof. Suchtreatment comprises the step of administering a therapeuticallyeffective amount of a compound of Formula (I) or (II) or apharmaceutically acceptable salt thereof, to said mammal, particularly ahuman Treatment can also comprise the step of administering atherapeutically effective amount of a pharmaceutical compositioncontaining a compound of Formula (I) or (II) or a pharmaceuticallyacceptable salt thereof, to said mammal, particularly a human.

As used herein, the term “effective amount” means that amount of a drugor pharmaceutical agent that will elicit the biological or medicalresponse of a tissue, system, animal, or human that is being sought, forinstance, by a researcher or clinician.

The term “therapeutically effective amount” means any amount which, ascompared to a corresponding subject who has not received such amount,results in improved treatment, healing, prevention, or amelioration of adisease, disorder, or side effect, or a decrease in the rate ofadvancement of a disease or disorder. The term also includes within itsscope amounts effective to enhance normal physiological function. Foruse in therapy, therapeutically effective amounts of a compound ofFormula (I) or (II), as well as salts thereof, may be administered asthe raw chemical. Additionally, the active ingredient may be presentedas a pharmaceutical composition. While it is possible that, for use intherapy, a therapeutically effective amount of a compound of Formula (I)or (II) or a pharmaceutically acceptable salt thereof, may beadministered as the raw chemical, it is typically presented as theactive ingredient of a pharmaceutical composition or formulation.

The precise therapeutically effective amount of a compound or saltthereof of the invention will depend on a number of factors, including,but not limited to, the age and weight of the subject (patient) beingtreated, the precise disorder requiring treatment and its severity, thenature of the pharmaceutical formulation/composition, and route ofadministration, and will ultimately be at the discretion of theattending physician or veterinarian. Typically, a compound of Formula(I) or (II) or a pharmaceutically acceptable salt thereof, will be givenfor the treatment in the range of about 0.1 to 100 mg/kg body weight ofrecipient (patient, mammal) per day and more usually in the range of 0.1to 10 mg/kg body weight per day. Acceptable daily dosages may be fromabout 0.1 to about 1000 mg/day, and preferably from about 1 to about 100mg/day. This amount may be given in a single dose per day or in a number(such as two, three, four, five, or more) of sub-doses per day such thatthe total daily dose is the same. An effective amount of a salt thereofmay be determined as a proportion of the effective amount of thecompound of Formula (I) or (II) per se. Similar dosages should beappropriate for treatment of the other conditions referred herein fortreatment. In general, determination of appropriate dosing can bereadily arrived at by one skilled in medicine or the pharmacy art.

The compounds of the invention may be used alone or in combination withone or more other therapeutic agents. Accordingly the present inventionprovides a combination comprising a compound of Formula (I) or apharmaceutically acceptable salt thereof and one or more othertherapeutic agents. Such combinations may be presented individually(wherein each active is in separate composition) or the actives arepresented in a combined composition.

The instant compounds can be combined with or co-administered with othertherapeutic agents, particularly agents that may enhance the activity ortime of disposition of the compounds. Combination therapies according tothe invention comprise the administration of at least one compound ofthe invention and the use of at least one other treatment method. In oneembodiment, combination therapies according to the invention comprisethe administration of at least one compound of the invention andsurgical therapy. In one embodiment, combination therapies according tothe invention comprise the administration of at least one compound ofthe invention and radiotherapy. In one embodiment, combination therapiesaccording to the invention comprise the administration of at least onecompound of the invention and at least one supportive care agent (e.g.,at least one anti-emetic agent). In one embodiment, combinationtherapies according to the present invention comprise the administrationof at least one compound of the invention and at least one otherchemotherapeutic agent. In one particular embodiment, the inventioncomprises the administration of at least one compound of the inventionand at least one anti-neoplastic agent. In yet another embodiment, theinvention comprises a therapeutic regimen where the RET inhibitors ofthis disclosure are not in and of themselves active or significantlyactive, but when combined with another therapy, which may or may not beactive as a standalone therapy, the combination provides a usefultherapeutic outcome.

By the term “co-administering” and derivatives thereof as used hereinrefers to either simultaneous administration or any manner of separatesequential administration of a RET inhibiting compound, as describedherein, and a further active ingredient or ingredients, particularlythose known to be useful in the treatment of cancer, includingchemotherapy and radiation treatment. The term further active ingredientor ingredients, as used herein, includes any compound or therapeuticagent known to or that demonstrates advantageous properties whenadministered to a patient in need of treatment for cancer. Preferably,if the administration is not simultaneous, the compounds areadministered in a close time proximity to each other. Furthermore, itdoes not matter if the compounds are administered in the same dosageform, e.g. one compound may be administered topically and anothercompound may be administered orally.

Typically, any anti-neoplastic agent that has activity versus asusceptible tumor being treated may be co-administered in the treatmentof specified cancers in the present invention. Examples of such agentscan be found in Cancer Principles and Practice of Oncology by V. T.Devita and S. Hellman (editors), 6^(th) edition (Feb. 15, 2001),Lippincott Williams & Wilkins Publishers. A person of ordinary skill inthe art would be able to discern which combinations of agents would beuseful based on the particular characteristics of the drugs and thecancer involved. Typical anti-neoplastic agents useful in the presentinvention include, but are not limited to, anti-microtubule agents suchas diterpenoids and vinca alkaloids; platinum coordination complexes;alkylating agents such as nitrogen mustards, oxazaphosphorines,alkylsulfonates, nitrosoureas, and triazenes; antibiotic agents such asanthracyclins, actinomycins and bleomycins; topoisomerase II inhibitorssuch as epipodophyllotoxins; antimetabolites such as purine andpyrimidine analogues and anti-folate compounds; topoisomerase Iinhibitors such as camptothecins; hormones and hormonal analogues; DNAmethyltransferase inhibitors such as azacitidine and decitabine; signaltransduction pathway inhibitors; non-receptor tyrosine kinaseangiogenesis inhibitors; immunotherapeutic agents; proapoptotic agents;and cell cycle signaling inhibitors.

Typically, any chemotherapeutic agent that has activity against asusceptible neoplasm being treated may be utilized in combination withthe compounds the invention, provided that the particular agent isclinically compatible with therapy employing a compound of theinvention. Typical anti-neoplastic agents useful in the presentinvention include, but are not limited to: alkylating agents,anti-metabolites, antitumor antibiotics, antimitotic agents, nucleosideanalogues, topoisomerase I and II inhibitors, hormones and hormonalanalogues; retinoids, histone deacetylase inhibitors; signaltransduction pathway inhibitors including inhibitors of cell growth orgrowth factor function, angiogenesis inhibitors, and serine/threonine orother kinase inhibitors; cyclin dependent kinase inhibitors; antisensetherapies and immunotherapeutic agents, including monoclonals, vaccinesor other biological agents.

Nucleoside analogues are those compounds which are converted todeoxynucleotide triphosphates and incorporated into replicating DNA inplace of cytosine. DNA methyltransferases become covalently bound to themodified bases resulting in an inactive enzyme and reduced DNAmethylation. Examples of nucleoside analogues include azacitidine anddecitabine which are used for the treatment of myelodysplastic disorder.Histone deacetylase (HDAC) inhibitors include vorinostat, for thetreatment of cutaneous T-cell lymphoma. HDACs modify chromatin throughthe deacetylation of histones. In addition, they have a variety ofsubstrates including numerous transcription factors and signalingmolecules. Other HDAC inhibitors are in development.

Signal transduction pathway inhibitors are those inhibitors which blockor inhibit a chemical process which evokes an intracellular change. Asused herein this change is cell proliferation or differentiation orsurvival. Signal transduction pathway inhibitors useful in the presentinvention include, but are not limited to, inhibitors of receptortyrosine kinases, non-receptor tyrosine kinases, SH2/SH3 domainblockers, serine/threonine kinases, phosphatidyl inositol-3-OH kinases,myoinositol signaling, and Ras oncogenes. Signal transduction pathwayinhibitors may be employed in combination with the compounds of theinvention in the compositions and methods described above.

Receptor kinase angiogenesis inhibitors may also find use in the presentinvention. Inhibitors of angiogenesis related to VEGFR and TIE-2 arediscussed above in regard to signal transduction inhibitors (both arereceptor tyrosine kinases). Other inhibitors may be used in combinationwith the compounds of the invention. For example, anti-VEGF antibodies,which do not recognize VEGFR (the receptor tyrosine kinase), but bind tothe ligand; small molecule inhibitors of integrin (alpha_(v) beta₃) thatinhibit angiogenesis; endostatin and angiostatin (non-RTK) may alsoprove useful in combination with the compounds of the invention. Oneexample of a VEGFR antibody is bevacizumab (AVASTIN®).

Several inhibitors of growth factor receptors are under development andinclude ligand antagonists, antibodies, tyrosine kinase inhibitors,anti-sense oligonucleotides and aptamers. Any of these growth factorreceptor inhibitors may be employed in combination with the compounds ofthe invention in any of the compositions and methods/uses describedherein. Trastuzumab (Herceptin®) is an example of an anti-erbB2 antibodyinhibitor of growth factor function. One example of an anti-erbB1antibody inhibitor of growth factor function is cetuximab (Erbitux™,C225). Bevacizumab (Avastin®) is an example of a monoclonal antibodydirected against VEGFR. Examples of small molecule inhibitors ofepidermal growth factor receptors include but are not limited tolapatinib (Tykerb®) and erlotinib (TARCEVA®). Imatinib mesylate(GLEEVEC®) is one example of a PDGFR inhibitor. Examples of VEGFRinhibitors include pazopanib (Votrient®), ZD6474, AZD2171, PTK787,sunitinib and sorafenib.

Anti-microtubule or anti-mitotic agents are phase specific agents activeagainst the microtubules of tumor cells during M or the mitosis phase ofthe cell cycle. Examples of anti-microtubule agents include, but are notlimited to, diterpenoids and vinca alkaloids.

Diterpenoids, which are derived from natural sources, are phase specificanti-cancer agents that operate at the G₂/M phases of the cell cycle. Itis believed that the diterpenoids stabilize the β-tubulin subunit of themicrotubules, by binding with this protein. Disassembly of the proteinappears then to be inhibited with mitosis being arrested and cell deathfollowing. Examples of diterpenoids include, but are not limited to,paclitaxel and its analog docetaxel.

Paclitaxel, 5β,20-epoxy-1,2α,4,7β,10β,13α-hexa-hydroxytax-11-en-9-one4,10-diacetate 2-benzoate 13-ester with(2R,3S)—N-benzoyl-3-phenylisoserine; is a natural diterpene productisolated from the Pacific yew tree Taxus brevifolia and is commerciallyavailable as an injectable solution TAXOL®. It is a member of the taxanefamily of terpenes. It was first isolated in 1971 by Wani et al. J. Am.Chem, Soc., 93:2325 (1971), who characterized its structure by chemicaland X-ray crystallographic methods. One mechanism for its activityrelates to paclitaxel's capacity to bind tubulin, thereby inhibitingcancer cell growth. Schiff et al., Proc. Natl, Acad, Sci. USA,77:1561-1565 (1980); Schiff et al., Nature, 277:665-667 (1979); Kumar,J. Biol, Chem, 256: 10435-10441 (1981). For a review of synthesis andanticancer activity of some paclitaxel derivatives see: D. G. I.Kingston et al., Studies in Organic Chemistry vol. 26, entitled “Newtrends in Natural Products Chemistry 1986”, Attaur-Rahman, P. W. LeQuesne, Eds. (Elsevier, Amsterdam, 1986) pp 219-235.

Paclitaxel has been approved for clinical use in the treatment ofrefractory ovarian cancer in the United States (Markman et al., YaleJournal of Biology and Medicine, 64:583, 1991; McGuire et al., Ann. Int.Med., 111:273, 1989) and for the treatment of breast cancer (Holmes etal., J. Nat. Cancer Inst., 83:1797, 1991). It is a potential candidatefor treatment of neoplasms in the skin (Einzig et. al., Proc. Am. Soc.Clin. Oncol., 20:46) and head and neck carcinomas (Forastire et. al.,Sem. Oncol., 20:56, 1990). The compound also shows potential for thetreatment of polycystic kidney disease (Woo et. al., Nature, 368:750,1994), lung cancer and malaria. Treatment of patients with paclitaxelresults in bone marrow suppression (multiple cell lineages, Ignoff, R.J. et. al, Cancer Chemotherapy Pocket Guide, 1998) related to theduration of dosing above a threshold concentration (50 nM) (Kearns, C.M. et. al., Seminars in Oncology, 3(6) p. 16-23, 1995).

Docetaxel, (2R,3S)—N-carboxy-3-phenylisoserine N-tert-butyl ester,13-ester with 5β-20-epoxy-1,2α,4,7β,10β,13α-hexahydroxytax-11-en-9-one4-acetate 2-benzoate, trihydrate; is commercially available as aninjectable solution as TAXOTERE®. Docetaxel is indicated for thetreatment of breast cancer. Docetaxel is a semisynthetic derivative ofpaclitaxel q.v., prepared using a natural precursor,10-deacetyl-baccatin III, extracted from the needle of the European Yewtree. The dose limiting toxicity of docetaxel is neutropenia.

Vinca alkaloids are phase specific anti-neoplastic agents derived fromthe periwinkle plant. Vinca alkaloids act at the M phase (mitosis) ofthe cell cycle by binding specifically to tubulin. Consequently, thebound tubulin molecule is unable to polymerize into microtubules.Mitosis is believed to be arrested in metaphase with cell deathfollowing. Examples of vinca alkaloids include, but are not limited to,vinblastine, vincristine, and vinorelbine.

Vinblastine, vincaleukoblastine sulfate, is commercially available asVELBAN® as an injectable solution. Although, it has possible indicationas a second line therapy of various solid tumors, it is primarilyindicated in the treatment of testicular cancer and various lymphomasincluding Hodgkin's Disease; and lymphocytic and histiocytic lymphomas.Myelosuppression is the dose limiting side effect of vinblastine.

Vincristine, vincaleukoblastine, 22-oxo-, sulfate, is commerciallyavailable as ONCOVIN® as an injectable solution. Vincristine isindicated for the treatment of acute leukemias and has also found use intreatment regimens for Hodgkin's and non-Hodgkin's malignant lymphomas.Alopecia and neurologic effects are the most common side effect ofvincristine and to a lesser extent myelosupression and gastrointestinalmucositis effects occur.

Vinorelbine, 3′,4′-didehydro-4′-deoxy-C′-norvincaleukoblastine[R—(R*,R*)-2,3-dihydroxybutanedioate (1:2)(salt)], commerciallyavailable as an injectable solution of vinorelbine tartrate(NAVELBINE®), is a semisynthetic vinca alkaloid. Vinorelbine isindicated as a single agent or in combination with otherchemotherapeutic agents, such as cisplatin, in the treatment of varioussolid tumors, particularly non-small cell lung, advanced breast, andhormone refractory prostate cancers. Myelosuppression is the most commondose limiting side effect of vinorelbine.

Platinum coordination complexes are non-phase specific anti-canceragents, which are interactive with DNA. The platinum complexes entertumor cells, undergo aquation and form intra- and interstrand crosslinkswith DNA causing adverse biological effects to the tumor. Examples ofplatinum coordination complexes include, but are not limited to,cisplatin and carboplatin.

Cisplatin, cis-diamminedichloroplatinum, is commercially available asPLATINOL® as an injectable solution. Cisplatin is primarily indicated inthe treatment of metastatic testicular and ovarian cancer and advancedbladder cancer. The primary dose limiting side effects of cisplatin arenephrotoxicity, which may be controlled by hydration and diuresis, andototoxicity.

Carboplatin, platinum, diammine[1,1-cyclobutane-dicarboxylate(2-)-O,O′], is commercially available asPARAPLATIN® as an injectable solution. Carboplatin is primarilyindicated in the first and second line treatment of advanced ovariancarcinoma. Bone marrow suppression is the dose limiting toxicity ofcarboplatin.

Alkylating agents are non-phase anti-cancer specific agents and strongelectrophiles. Typically, alkylating agents form covalent linkages, byalkylation, to DNA through nucleophilic moieties of the DNA moleculesuch as phosphate, amino, sulfhydryl, hydroxyl, carboxyl, and imidazolegroups. Such alkylation disrupts nucleic acid function leading to celldeath. Examples of alkylating agents include, but are not limited to,nitrogen mustards such as cyclophosphamide, melphalan, and chlorambucil;alkyl sulfonates such as busulfan; nitrosoureas such as carmustine; andtriazenes such as dacarbazine.

Cyclophosphamide,2-[bis(2-chloroethyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorine 2-oxidemonohydrate, is commercially available as an injectable solution ortablets as CYTOXAN®. Cyclophosphamide is indicated as a single agent orin combination with other chemotherapeutic agents, in the treatment ofmalignant lymphomas, multiple myeloma, and leukemias. Alopecia, nausea,vomiting and leukopenia are the most common dose limiting side effectsof cyclophosphamide.

Melphalan, 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commerciallyavailable as an injectable solution or tablets as ALKERAN®. Melphalan isindicated for the palliative treatment of multiple myeloma andnon-resectable epithelial carcinoma of the ovary. Bone marrowsuppression is the most common dose limiting side effect of melphalan.

Chlorambucil, 4-[bis(2-chloroethyl)amino]benzenebutanoic acid, iscommercially available as LEUKERAN® tablets. Chlorambucil is indicatedfor the palliative treatment of chronic lymphatic leukemia, andmalignant lymphomas such as lymphosarcoma, giant follicular lymphoma,and Hodgkin's disease. Bone marrow suppression is the most common doselimiting side effect of chlorambucil.

Busulfan, 1,4-butanediol dimethanesulfonate, is commercially availableas MYLERAN® TABLETS. Busulfan is indicated for the palliative treatmentof chronic myelogenous leukemia. Bone marrow suppression is the mostcommon dose limiting side effects of busulfan.

Carmustine, 1,3-[bis(2-chloroethyl)-1-nitrosourea, is commerciallyavailable as single vials of lyophilized material as BiCNU®. Carmustineis indicated for the palliative treatment as a single agent or incombination with other agents for brain tumors, multiple myeloma,Hodgkin's disease, and non-Hodgkin's lymphomas. Delayed myelosuppressionis the most common dose limiting side effects of carmustine.

Dacarbazine, 5-(3,3-dimethyl-1-triazeno)-imidazole-4-carboxamide, iscommercially available as single vials of material as DTIC-Dome®.Dacarbazine is indicated for the treatment of metastatic malignantmelanoma and in combination with other agents for the second linetreatment of Hodgkin's Disease. Nausea, vomiting, and anorexia are themost common dose limiting side effects of dacarbazine.

Antibiotic anti-neoplastics are non-phase specific agents, which bind orintercalate with DNA. Typically, such action results in stable DNAcomplexes or strand breakage, which disrupts ordinary function of thenucleic acids leading to cell death. Examples of antibioticanti-neoplastic agents include, but are not limited to, actinomycinssuch as dactinomycin, anthrocyclins such as daunorubicin anddoxorubicin; and bleomycins.

Dactinomycin, also known as Actinomycin D, is commercially available ininjectable form as COSMEGEN®. Dactinomycin is indicated for thetreatment of Wilm's tumor and rhabdomyosarcoma. Nausea, vomiting, andanorexia are the most common dose limiting side effects of dactinomycin.

Daunorubicin,(8S-cis-)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)-oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12naphthacenedione hydrochloride, is commercially available as a liposomalinjectable form as DAUNOXOME® or as an injectable as CERUBIDINE®.Daunorubicin is indicated for remission induction in the treatment ofacute nonlymphocytic leukemia and advanced HIV associated Kaposi'ssarcoma. Myelosuppression is the most common dose limiting side effectof daunorubicin.

Doxorubicin,(8S,10S)-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-8-glycoloyl,7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12naphthacenedione hydrochloride, is commercially available as aninjectable form as RUBEX® or ADRIAMYCIN RDF®. Doxorubicin is primarilyindicated for the treatment of acute lymphoblastic leukemia and acutemyeloblastic leukemia, but is also a useful component in the treatmentof some solid tumors and lymphomas. Myelosuppression is the most commondose limiting side effect of doxorubicin.

Bleomycin, a mixture of cytotoxic glycopeptide antibiotics isolated froma strain of Streptomyces verticillus, is commercially available asBLENOXANE®. Bleomycin is indicated as a palliative treatment, as asingle agent or in combination with other agents, of squamous cellcarcinoma, lymphomas, and testicular carcinomas. Pulmonary and cutaneoustoxicities are the most common dose limiting side effects of bleomycin.

Topoisomerase II inhibitors include, but are not limited to,epipodophyllotoxins.

Epipodophyllotoxins are phase specific anti-neoplastic agents derivedfrom the mandrake plant. Epipodophyllotoxins typically affect cells inthe S and G₂ phases of the cell cycle by forming a ternary complex withtopoisomerase II and DNA causing DNA strand breaks. The strand breaksaccumulate and cell death follows. Examples of epipodophyllotoxinsinclude, but are not limited to, etoposide and teniposide.

Etoposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-ethylidene-β-D-glucopyranoside], is commercially availableas an injectable solution or capsules as VePESID® and is commonly knownas VP-16. Etoposide is indicated as a single agent or in combinationwith other chemotherapy agents in the treatment of testicular andnon-small cell lung cancers. Myelosuppression is the most common sideeffect of etoposide. The incidence of leukopenialeukopenia tends to bemore severe than thrombocytopenia.

Teniposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-thenylidene-β-D-glucopyranoside], is commercially availableas an injectable solution as VUMON® and is commonly known as VM-26.Teniposide is indicated as a single agent or in combination with otherchemotherapy agents in the treatment of acute leukemia in children.Myelosuppression is the most common dose limiting side effect ofteniposide. Teniposide can induce both leukopenialeukopenia andthrombocytopenia.

Antimetabolite neoplastic agents are phase specific anti-neoplasticagents that act at S phase (DNA synthesis) of the cell cycle byinhibiting DNA synthesis or by inhibiting purine or pyrimidine basesynthesis and thereby limiting DNA synthesis. Consequently, S phase doesnot proceed and cell death follows. Examples of antimetaboliteanti-neoplastic agents include, but are not limited to, fluorouracil,methotrexate, cytarabine, mecaptopurine, thioguanine, and gemcitabine.

5-fluorouracil, 5-fluoro-2,4-(1H,3H)pyrimidinedione, is commerciallyavailable as fluorouracil. Administration of 5-fluorouracil leads toinhibition of thymidylate synthesis and is also incorporated into bothRNA and DNA. The result typically is cell death. 5-fluorouracil isindicated as a single agent or in combination with other chemotherapyagents in the treatment of carcinomas of the breast, colon, rectum,stomach and pancreas. Myelosuppression and mucositis are dose limitingside effects of 5-fluorouracil. Other fluoropyrimidine analogs include5-fluoro deoxyuridine (floxuridine) and 5-fluorodeoxyuridinemonophosphate.

Cytarabine, 4-amino-1-β-D-arabinofuranosyl-2 (1H)-pyrimidinone, iscommercially available as CYTOSAR-U® and is commonly known as Ara-C. Itis believed that cytarabine exhibits cell phase specificity at S-phaseby inhibiting DNA chain elongation by terminal incorporation ofcytarabine into the growing DNA chain. Cytarabine is indicated as asingle agent or in combination with other chemotherapy agents in thetreatment of acute leukemia. Other cytidine analogs include5-azacytidine and 2′,2′-difluorodeoxycytidine (gemcitabine). Cytarabineinduces leukopenialeukopenia, thrombocytopenia, and mucositis.

Mercaptopurine, 1,7-dihydro-6H-purine-6-thione monohydrate, iscommercially available as PURINETHOL®. Mercaptopurine exhibits cellphase specificity at S-phase by inhibiting DNA synthesis by an as of yetunspecified mechanism. Mercaptopurine is indicated as a single agent orin combination with other chemotherapy agents in the treatment of acuteleukemia. Myelosuppression and gastrointestinal mucositis are expectedside effects of mercaptopurine at high doses. A useful mercaptopurineanalog is azathioprine.

Thioguanine, 2-amino-1,7-dihydro-6H-purine-6-thione, is commerciallyavailable as TABLOID®. Thioguanine exhibits cell phase specificity atS-phase by inhibiting DNA synthesis by an as of yet unspecifiedmechanism. Thioguanine is indicated as a single agent or in combinationwith other chemotherapy agents in the treatment of acute leukemia.Myelosuppression, including leukopenialeukopenia, thrombocytopenia, andanemia, is the most common dose limiting side effect of thioguanineadministration. However, gastrointestinal side effects occur and can bedose limiting. Other purine analogs include pentostatin,erythrohydroxynonyladenine, fludarabine phosphate, and cladribine.

Gemcitabine, 2′-deoxy-2′,2′-difluorocytidine monohydrochloride(β-isomer), is commercially available as GEMZAR®. Gemcitabine exhibitscell phase specificity at S-phase and by blocking progression of cellsthrough the G1/S boundary. Gemcitabine is indicated in combination withcisplatin in the treatment of locally advanced non-small cell lungcancer and alone in the treatment of locally advanced pancreatic cancer.Myelosuppression, including leukopenialeukopenia, thrombocytopenia, andanemia, is the most common dose limiting side effect of gemcitabineadministration.

Methotrexate,N-[4[[(2,4-diamino-6-pteridinyl)methyl]methylamino]benzoyl]-L-glutamicacid, is commercially available as methotrexate sodium. Methotrexateexhibits cell phase effects specifically at S-phase by inhibiting DNAsynthesis, repair and/or replication through the inhibition ofdyhydrofolic acid reductase which is required for synthesis of purinenucleotides and thymidylate. Methotrexate is indicated as a single agentor in combination with other chemotherapy agents in the treatment ofchoriocarcinoma, meningeal leukemia, non-Hodgkin's lymphoma, andcarcinomas of the breast, head, neck, ovary and bladder.Myelosuppression (leukopenia, thrombocytopenia, and anemia) andmucositis are expected side effect of methotrexate administration.

Camptothecins, including, camptothecin and camptothecin derivatives areavailable or under development as Topoisomerase I inhibitors.Camptothecins cytotoxic activity is believed to be related to itsTopoisomerase I inhibitory activity. Examples of camptothecins include,but are not limited to irinotecan, topotecan, and the various opticalforms of7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20-camptothecindescribed below.

Irinotecan HCl,(4S)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)carbonyloxy]-1H-pyrano[3′,4′,6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dionehydrochloride, is commercially available as the injectable solutionCAMPTOSAR®.

Irinotecan is a derivative of camptothecin which binds, along with itsactive metabolite SN-38, to the topoisomerase I-DNA complex. It isbelieved that cytotoxicity occurs as a result of irreparable doublestrand breaks caused by interaction of the topoisomerase I:DNA:irintecanor SN-38 ternary complex with replication enzymes. Irinotecan isindicated for treatment of metastatic cancer of the colon or rectum. Thedose limiting side effects of irinotecan HCl are myelosuppression,including neutropenia, and GI effects, including diarrhea.

Topotecan HCl,(S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3′,4′,6,7]indolizino[1,2-b]quinoline-3,14-(4H,12H)-dionemonohydrochloride, is commercially available as the injectable solutionHYCAMTIN®. Topotecan is a derivative of camptothecin which binds to thetopoisomerase I-DNA complex and prevents religation of singles strandbreaks caused by Topoisomerase I in response to torsional strain of theDNA molecule. Topotecan is indicated for second line treatment ofmetastatic carcinoma of the ovary and small cell lung cancer. The doselimiting side effect of topotecan HCl is myelosuppression, primarilyneutropenia.

Experimentals

The following examples illustrate the invention. These examples are notintended to limit the scope of the present invention, but rather toprovide guidance to the skilled artisan to prepare and use thecompounds, compositions, and methods of the present invention. Whileparticular embodiments of the present invention are described, theskilled artisan will appreciate that various changes and modificationscan be made without departing from the spirit and scope of theinvention. Unless otherwise noted, reagents are commercially availableor are prepared according to procedures in the literature. The symbolsand conventions used in the descriptions of processes, schemes, andexamples are consistent with those used in the contemporary scientificliterature, for example, the Journal of the American Chemical Society orthe Journal of Biological Chemistry.

In the Examples:

Chemical shifts are expressed in parts per million (ppm) units. Couplingconstants (J) are in units of hertz (Hz). Splitting patterns describeapparent multiplicities and are designated as s (singlet), d (doublet),t (triplet), q (quartet), dd (double doublet), dt (double triplet), dq(double quartet), m (multiplet), br (broad).

Flash column chromatography was performed on silica gel.

The naming program used was ChemBioDraw® Ultra 12.0.

Abbreviations

-   18-crown-6 1,4,7,10,13,16-hexaoxacyclooctadecane-   CDCl₃ chloroform-d-   CD₃OD methanol-d₄-   Cs₂CO₃ cesium carbonate-   d day(s)-   DCM dichloromethane-   DMF N,N-dimethylformamide-   EA ethyl acetate-   ES-LCMS electrospray liquid chromatography-mass spectrometry-   Et₃N triethylamine-   EtOH ethanol-   g gram(s)-   h hour(s)-   H₂ hydrogen gas-   HCl hydrochloric acid-   H₂O water-   HPLC high performance liquid chromatography-   H₂SO₄ sulfuric acid-   K₂CO₃ potassium carbonate-   KOAc potassium acetate-   KOH potassium hydroxide-   LCMS liquid chromatography-mass spectrometry-   LiOH.H₂O lithium hydroxide hydrate-   MeCN acetonitrile-   MeOH methanol-   mg milligram(s)-   MgSO₄ magnesium sulfate-   min minute(s)-   mL milliliter(s)-   mmol millimole(s)-   N₂ nitrogen gas-   NaCN sodium cyanide-   NaHCO₃ sodium bicarbonate-   NaOH sodium hydroxide-   Na₂SO₄ sodium sulphate-   NBS N-bromosuccinimide-   NH₄OH ammonium hydroxide-   NMR nuclear magnetic resonance-   Pd/C palladium on carbon-   PdCl₂(dppf)    1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II)-   PE petroleum ether-   PMB p-methoxybenzyl-   rt room temperature-   SOCl₂ thionyl chloride-   TBME tert-butyl methyl ether-   TFA trifluoroacetic acid-   TLC thin layer chromotrography-   T₃P® propylphosphonic anhydride

Preparation of Intermediates Intermediate 1:2-(4-(4-Ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)aceticacid

Step 1: 2-Chloro-4-ethoxypyridine

To a mixture of 2-chloro-4-nitropyridine (170 g, 1070 mmol) in THF (2 L)was added sodium ethanolate (109.45 g, 1610 mmol) slowly at 0° C. Themixture was stirred at 25° C. for 12 h. LCMS and TLC analysis(PE/EA=5:1, R_(f)=0.6) showed the reaction was finished. The mixture wasfiltered, and most of the filtrate solvent was removed by reducedpressure. The mixture was quenched with water and extracted with EA, theorganic layer was washed with brine, and then concentrated. Another sixbatches were prepared following the same procedure to give2-chloro-4-ethoxypyridine (1100 g, 7.01 mol, 92.4%): ¹H NMR (400 MHz,CD₃OD) δ 8.15 (d, J=6.0 Hz, 1H), 6.99 (d, J=2.0 Hz, 1H), 6.91-6.89 (m,1H), 4.16-4.14 (m, 2H), 1.41-1.38 (m, 3H); ES-LCMS m/z: 158.1 (M+H).

Step 2: 5-Bromo-2-chloro-4-ethoxypyridine

2-Chloro-4-ethoxypyridine (100 g, 634.5 mmol) was added to H₂SO₄ (500mL) slowly. NBS (124.2 g, 698.0 mmol) was then added to the abovereaction mixture at rt. The mixture was stirred at 80° C. for 3 h. TLCanalysis (PE/EA=10:1, R_(f)=0.5) showed the reaction was finished. Thereaction mixture was poured into ice-water (2000 mL), extracted with EA,and then concentrated. Another ten batches were prepared following thesame procedure. The combined crude product was purified by flash columnchromatography to give 5-bromo-2-chloro-4-ethoxypyridine (670 g, 2.84mol, 40.0%): ¹H NMR (400 MHz, CD₃OD): δ 8.31 (s, 1H), 7.14 (s, 1H),4.32-4.10 (m, 2H), 1.58-1.35 (m, 3H); ES-LCMS m/z: 236.0, 238.0 (M,M+2H).

Step 3: 5-Bromo-4-ethoxy-2((4-methoxybenzyl)oxy)pyridine

To a mixture of 5-bromo-2-chloro-4-ethoxypyridine (75 g, 317.1 mmol) intoluene (500 mL) was added (4-methoxyphenyl)methanol (52.6 g, 380.6mmol), KOH (35.6 g, 634.3 mmol) and 18-crown-6 (8.4 g, 31.2 mmol) at rt.The reaction mixture was stirred at 120° C. for 2 h. The mixture wasextracted with TBME, washed with brine, and concentrated. Another eightbatches were prepared following the same procedure. The combined crudeproduct was purified by flash column chromatography (PE/EA=10:1,R_(f)=0.5) to give 5-bromo-4-ethoxy-2-((4-methoxybenzyl)oxy)pyridine(650 g, 1.99 mol, 70.0%): ¹H NMR (400 MHz, CD₃OD) δ 8.05 (s, 1H), 7.33(d, J=8.6 Hz, 2H), 6.90-6.84 (m, 2H), 6.38 (s, 1H), 5.20 (s, 2H),4.16-4.05 (m, 2H), 3.77 (s, 3H), 1.43 (q, J=6.8 Hz, 3H); ES-LCMS m/z:338.3 (M+2H).

Step 4: 2-(4-Bromo-2-fluorophenyl)acetonitrile

To a solution of 4-bromo-1-(bromomethyl)-2-fluorobenzene (500 g, 1.87mol) in EtOH (2.2 L) stirred under N₂ at 20° C. was added NaCN (93 g,1.90 mmol) in one charge. The reaction mixture was stirred at 60° C. for12 h. Then the solution was concentrated and distributed between DCM(2000 mL) and saturated NaHCO₃ solution (1800 mL). Another batch wasprepared following the same procedure. Then the two batches werecombined. The combined organic extract was washed with brine, dried overMgSO₄, filtered and concentrated to provide2-(4-bromo-2-fluorophenyl)acetonitrile (794 g, 99%): ¹H NMR (400 MHz,CDCl₃) δ 7.38-7.27 (m, 3H), 3.72 (s, 2H).

Step 5: 2-(4-Bromo-2-fluorophenyl)acetic acid

To a solution of 2-(4-bromo-2-fluorophenyl)acetonitrile (397 g, 1.82mol) in MeOH (500 mL) stirred under N₂ at 20° C. was added NaOH (2.22 L,2.5M, 5.56 mol) solution in one charge. The reaction mixture was stirredat 80° C. for 5 h. Then the solution was concentrated and neutralizedwith conc. HCl to pH=5 with stirring. Then the solution was extractedwith EA (1.5 L×2). Another two batches were prepared following the sameprocedure. Then the three batches were combined. The combined organicextract was washed with brine, dried over Na₂SO₄, filtered andconcentrated in vacuo to give the pure 2-(4-bromo-2-fluorophenyl)aceticacid (1200 g, 92%): TLC (PE/EA=5:1, R_(f)=0.2); ¹H NMR (400 MHz, CDCl₃)δ 7.24 (br. s., 1H), 7.12 (t, J=7.9 Hz, 1H), 3.65 (s, 2H).

Step 6: Methyl 2-(4-bromo-2-fluorophenyl)acetate

To a solution of 2-(4-bromo-2-fluorophenyl)acetic acid (260 g, 1.13 mol)in MeOH (2 L) was added H₂SO₄ (30 mL) at rt. The solution was heated toreflux overnight. Then the solvent was concentrated and the residue wasdistributed between EA and saturated NaHCO₃ solution. The organicextract was washed with brine, dried over Na₂SO₄, filtered andconcentrated. Another batch was prepared following the same procedure.Then the two batches were combined to provide methyl2-(4-bromo-2-fluorophenyl)acetate (520 g, 94%). TLC (PE/EA=10:1,R_(f)=0.7). ¹H NMR (400 MHz, CDCl₃) δ 7.25-7.20 (m, 2H), 7.14 (t, J=8.0Hz, 1H), 3.70 (s, 3H), 3.62 (s, 2H).

Step 7: Methyl2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetate

To a solution of methyl 2-(4-bromo-2-fluorophenyl)acetate (260 g, 1.05mol) and 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane)(320 g, 1.26 mol) in 1,4-dioxane (2 L) was added KOAc (206 g, 2.10 mol)and PdCl₂(dppf) (23 g, 0.03 mol) at rt. The solution was heated toreflux for 4 h under N₂. Then the solution was filtered and the filtratewas concentrated in vacuo to give the crude product. Another batch wasprepared following the same procedure. Then the two batches werecombined and purified by flash column chromatography (PE/EA=30:1 to10:1). All fractions found to contain product by TLC (PE/EA=10:1,R_(f)=0.5) were combined and concentrated to yield methyl2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetate(560 g, 90%) as a light yellow oil: ¹H NMR (400 MHz, CDCl₃) δ 7.54 (d,J=7.5 Hz, 1H), 7.49 (d, J=10.0 Hz, 1H), 7.31-7.26 (m, 1H), 3.73 (s, 2H),1.34 (s, 12H), 1.27 (s, 3H); ES-LCMS m/z 295.2 (M+H).

Step 8: Methyl2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetate

To a solution of 5-bromo-4-ethoxy-2-((4-methoxybenzyl)oxy)pyridine (175g, 519 mmol) in 1,4-dioxane (1200 mL) and H₂O (300 mL) was added methyl2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetate(167 g, 569 mmol), PdCl₂(dppf) (25 g, 5.19 mmol) and Cs₂CO₃ (337 g, 1038mmol) under a N₂ atmosphere. The mixture was refluxed for 2 h. TLCanalysis (PE/EA=5:1, R_(f)=0.3) showed the reaction was finished. Themixture was extracted with EA/H₂O (2 L) to give the oil layer, which wasdried over Na₂SO₄, filtered, concentrated. Another two batches wereprepared following the same procedure. The combined crude product waspurified by flash column chromatography (PE/EA=5:1, R_(f)=0.3) to give5-bromo-4-ethoxy-2-((4-methoxybenzyl)oxy)pyridine (630 g, 1.48 mol,90.0%): ¹H NMR (400 MHz, CD₃OD) δ 7.94 (s, 1H), 7.36 (d, J=8.8 Hz, 2H),7.32-7.22 (m, 3H), 6.90 (d, J=8.8 Hz, 2H), 6.43 (s, 1H), 5.26 (s, 2H),4.11 (d, J=6.8 Hz, 2H), 3.78 (s, 3H), 3.72 (s, 2H), 3.70 (s, 3H), 1.36(t, J=7.0 Hz, 3H); ES-LCMS m/z: 426.1 (M+H).

Step 9:2-(4-(4-Ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)aceticacid

To a solution of methyl2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetate(210 g, 519 mmol) in THF (500 mL) was added LiOH.H₂O (52 g, 1230 mmol)in H₂O (700 mL). The mixture was stirred at 60° C. overnight. TLCanalysis (PE/EA=5:1, Rf=0.3) showed the reaction was finished. Themixture was concentrated, and adjusted with HCl (1 N) to pH=7. Anothertwo batches were prepared following the same procedure. Then thecombined crude product was filtered, the solid was washed with water anddried to give2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)aceticacid (550 g, 1.34 mol, 93.0%): ¹H NMR (400 MHz, CD₃OD): δ 7.94 (s, 1H),7.41-7.28 (m, 3H), 7.24 (d, J=9.5 Hz, 2H), 6.91 (d, J=8.6 Hz, 2H), 6.44(s, 1H), 5.26 (s, 2H), 4.11 (q, J=6.9 Hz, 2H), 3.78 (s, 3H), 3.67 (s,2H), 1.36 (t, J=7.0 Hz, 3H); ES-LCMS m/z: 412.1 (M+H).

Preparation of Compounds of the Invention Example 1N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide

Step 1: 3-Nitro-5-(trifluoromethyl)benzoic acid

To a mixture of 3-(trifluoromethyl)benzoic acid (5 g, 26.3 mmol) inH₂SO₄ (50 mL, 938 mmol) was added nitric acid (3.53 mL, 79 mmol). Themixture was stirred at 0° C. for 15 min and warmed to 90° C. over 1 h.Then the mixture was added to ice water dropwise. The mixture wasfiltered to give a white solid of 3-nitro-5-(trifluoromethyl)benzoicacid (5.2 g, 21.01 mmol, 80.0%): ¹H NMR (400 MHz, CD₃OD) δ 8.99 (s, 1H),8.72 (s, 1H), 8.61 (s, 1H).

Step 2: N-(2-(dimethylamino)ethyl)-3-nitro-5-(trifluoromethyl)benzamide

A mixture of 3-nitro-5-(trifluoromethyl)benzoic acid (3.3 g, 14.04 mmol)in SOCl₂ (50 mL, 685 mmol) and DMF (0.5 mL, 6.46 mmol) was stirred at75° C. for 2 h. Then the mixture was concentrated, and the acid chloridewas dissolved in DCM (50 mL). The mixture cooled to 0° C. and then wasadded Et₃N (2.348 mL, 16.84 mmol) followed byN¹,N¹-dimethylethane-1,2-diamine (0.740 mL, 15.44 mmol). The reactionwas stirred at 0° C. for 30 min and the mixture was then concentrated.The crude material was purified by flash column chromatography(DCM/MeOH=15:1). All fractions found to contain product by TLC(DCM/MeOH=15:1, R_(f)=0.3) were combined and concentrated to yield ayellow oil ofN-(2-(dimethylamino)ethyl)-3-nitro-5-(trifluoromethyl)benzamide (5 g,8.19 mmol, 58.4%): ¹H NMR (400 MHz, CD₃OD) δ 9.00 (s, 1H), 8.65 (s, 1H),8.61 (s, 1H), 3.86 (t, J=6.0 Hz, 2H), 3.42 (t, J=6.0 Hz, 2H), 2.99 (s,6H); ES-LCMS m/z 306.1 (M+H).

Step 3: 3-Amino-N-(2-(dimethylamino)ethyl)-5-(trifluoromethyl)benzamidedihydrochloride

To a mixture ofN-(2-(dimethylamino)ethyl)-3-nitro-5-(trifluoromethyl)benzamide (5 g,8.19 mmol) in MeOH (100 mL) was added Pd/C (500 mg, 10%). The mixturewas stirred at 10° C. for 16 h under a H₂ atmosphere. Then the mixturewas filtered and concentrated. The crude material was purified bypreparative HPLC (Column: ASB C18 150*25 mm; Mobile phase A: H₂O+0.1%HCl; Mobile phase B: MeCN; Flowrate: 25 mL/min; Gradient ProfileDescription: 5-35 (B %)) to yield an off-white solid of3-amino-N-(2-(dimethylamino)ethyl)-5-(trifluoromethyl)benzamidedihydrochloride (878.01 mg, 2.52 mmol, 30.8%): ¹H NMR (400 MHz, CD₃OD) δ8.28 (s, 1H), 8.19 (s, 1H), 7.86 (s, 1H), 3.84 (t, J=6.0 Hz, 2H), 3.47(t, J=6.0 Hz, 2H), 3.02 (s, 6H); ES-LCMS m/z 276.1 (M+H).

Step 4:N-(2-(dimethylamino)ethyl)-3-(3-(2-(5-ethoxy-6-((4-methoxybenzyl)oxy)pyridine-3-yl)-4-methylpyrimidin-5-yl)ureido)-5-(trifluoromethyl)benzamide

To a mixture of2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)aceticacid (100 mg, 0.243 mmol) in pyridine (3 mL) were added3-amino-N-(2-(dimethylamino)ethyl)-5-(trifluoromethyl)benzamide (66.9mg, 0.243 mmol) and T₃P® (0.8 mL, 0.243 mmol, 50% in EA). The mixturewas stirred at 10° C. for 16 h. The mixture was concentrated. The crudematerial was purified by preparative TLC (DCM/MeOH=10:1, R_(f)=0.3) toyield a yellow solid ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide(80 mg, 0.096 mmol, 39.4%): ¹H NMR (400 MHz, CD₃OD): δ 8.55 (d, J=4.4Hz, 1H), 8.43 (s, 1H), 8.06 (s, 1H), 7.96 (s, 1H), 7.89 (s, 1H),7.48-7.45 (m, 1H), 7.36 (d, J=8.8 Hz, 2H), 7.29 (s, 1H), 6.90 (d, J=8.8Hz, 2H), 6.45 (s, 1H), 5.26 (s, 2H), 4.12 (q, J=7.2 Hz, 2H), 3.84 (s,2H), 3.78 (s, 3H), 3.76 (t, J=2.4 Hz, 2H), 3.37 (t, J=5.6 Hz, 2H), 2.97(s, 6H), 1.36 (t, J=7.2 Hz, 3H); ES-LCMS m/z 669.1 (M+H).

Step 5:N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide

To a mixture ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide(80 mg, 0.120 mmol) in DCM (20 mL) was added TFA (2 mL, 26.0 mmol). Themixture was stirred at 25° C. for 2 h. Then the reaction wasconcentrated and basified with NH₄OH. The crude material was purified bypreparative HPLC (Instrument: Gilson GX281; Column: Gemini 150*25 mm*5um; Mobile phase A: H₂O (0.05% ammonia solution); Mobile phase B: MeCN;Gradient: 25-55 (B %); Flowrate: 25 mL/min; Run time: 10 min) to yieldan white solid ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide(27.71 mg, 0.050 mmol, 41.8%): TLC (DCM/MeOH=10:1, Rf=0.2) ¹H NMR (400MHz, CD₃OD): δ 8.26 (s, 1H), 8.20 (s, 1H), 7.90 (s, 1H), 7.45-7.39 (m,2H), 7.29-7.23 (m, 2H), 6.02 (s, 1H), 4.14 (q, J=7.2 Hz, 2H), 3.86 (s,2H), 3.57 (t, J=6.4 Hz, 2H), 2.63 (t, J=6.8 Hz, 2H), 2.36 (s, 6H), 1.40(t, J=7.2 Hz, 3H); ES-LCMS m/z 549.2 (M+H).

Example 2N-(3-(2-(dimethylamino)ethoxy)-5-(trifluoromethyl)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide

Step 1: N,N-dimethyl-2-(3-nitro-5-(trifluoromethyl)phenoxy)ethanamine

To a suspension of 1-fluoro-3-nitro-5-(trifluoromethyl)benzene (2 g,9.56 mmol) in DMF (15 mL) were added 2-(dimethylamino)ethanol (2.56 g,28.7 mmol) and K₂CO₃ (2.64 g, 19.13 mmol). The mixture was stirred at80° C. for 8 h. The mixture was cooled to rt. Then the solution wasconcentrated and distributed between EA (10 mL) and a saturated NaHCO₃solution (10 mL). The organic extract was washed with brine (10 mL),dried over MgSO₄, filtered, and concentrated. The crude material waspurified by flash column chromatography (PE/EA=1:1). All fractions foundto contain product by TLC (PE/EA=1:1, R_(f)=0.5) were combined andconcentrated to yield a light yellow solid ofN,N-dimethyl-2-(3-nitro-5-(trifluoromethyl)phenoxy)ethanamine (1.8 g,5.18 mmol, 54.1%): ¹H NMR (400 MHz, CDCl₃) δ 8.07 (s, 1H), 7.94 (s, 1H),7.50 (s, 1H), 4.18 (t, J=5.4 Hz, 2H), 2.79 (t, J=5.4 Hz, 2H), 2.36 (s,6H); ES-LCMS m/z 279.0 (M+H).

Step 2: 3-(2-(Dimethylamino)ethoxy)-5-(trifluoromethyl)aniline

To a suspension ofN,N-dimethyl-2-(3-nitro-5-(trifluoromethyl)phenoxy)ethanamine (1.8 g,6.47 mmol) in MeOH (20 mL) was added Pd/C (0.344 g, 10%). The mixturewas hydrogenated under a H₂ atmosphere at 15 Psi at 26° C. for 3 h. Thenthe solution was filtered and concentrated. The crude material waspurified by flash column chromatography (PE/EA=1:1). All fractions foundto contain product by TLC (PE/EA=1:1, R_(f)=0.4) were combined andconcentrated to yield a light yellow solid of3-(2-(dimethylamino)ethoxy)-5-(trifluoromethyl)aniline (1 g, 3.54 mmol,54.7%): ¹H NMR (400 MHz, CD₃OD) δ 6.53 (d, J=16.8 Hz, 2H), 6.38 (s, 1H),4.05 (t, J=5.6 Hz, 2H), 2.75-2.70 (m, 2H), 2.35-2.32 (m, 6H); ES-LCMSm/z 249.1 (M+H).

Step 3:N-(3-(2-(dimethylamino)ethoxy)-5-(trifluoromethyl)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide

To a solution of2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)aceticacid (400 mg, 0.972 mmol) and3-(2-(dimethylamino)ethoxy)-5-(trifluoromethyl)aniline (241 mg, 0.972mmol) in pyridine (8 mL) was added T₃P® (4 mL, 0.972 mmol) at 27° C.under a N₂ atmosphere. The mixture was stirred at 27° C. for 30 min.LCMS showed the reaction was completed. Then the mixture was put intoice (1 g). The mixture was concentrated to a residue that was purifiedby preparative TLC (DCM/MeOH=10:1, R_(f)=0.6) to yield a light yellowsolid ofN-(3-(2-(dimethylamino)ethoxy)-5-(trifluoromethyl)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide(300 mg, 0.421 mmol, 43.3%): ¹H NMR (400 MHz, CD₃OD) δ 7.96 (s, 1H),7.75-7.68 (m, 1H), 7.52 (s, 1H), 7.38 (d, J=8.4 Hz, 2H), 7.32-7.24 (m,3H), 7.05 (s, 1H), 6.92 (d, J=8.6 Hz, 2H), 6.45 (s, 1H), 5.27 (s, 2H),4.43-4.36 (m, 2H), 4.12 (q, J=6.8 Hz, 2H), 3.83 (s, 2H), 3.82-3.77 (m,3H), 3.60-3.51 (m, 2H), 2.93 (s, 6H), 1.37 (t, J=6.9 Hz, 3H); ES-LCMSm/z 642.2 (M+H).

Step 4:N-(3-(2-(dimethylamino)ethoxy)-5-(trifluoromethyl)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide

To a suspension ofN-(3-(2-(dimethylamino)ethoxy)-5-(trifluoromethyl)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide(300 mg, 0.468 mmol) in MeOH (10 mL) was added Pd/C (49.8 mg, 10%). Themixture was hydrogenated under a H₂ atmosphere at 25° C. for 3 h. Thenthe reaction was filtered and concentrated to a residue that waspurified by preparative HPLC (Instrument: Gilson GX281/Column: Gemini150*25 mm*5 um/Mobile phase A: H₂O (0.05% ammonia solution)/Mobile phaseB: MeCN/Gradient: 40-70 (B %)/Flowrate: 25 mL/min/Run time: 10 min) toyield a white solid ofN-(3-(2-(dimethylamino)ethoxy)-5-(trifluoromethyl)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide(54.34 mg, 0.104 mmol, 22.2%): ¹H NMR (400 MHz, CD₃OD) δ 7.52 (d, J=9.9Hz, 2H), 7.43-7.31 (m, 2H), 7.27-7.17 (m, 2H), 6.95 (s, 1H), 5.99 (s,1H), 4.21-4.04 (m, 4H), 3.80 (s, 2H), 2.79 (t, J=5.3 Hz, 2H), 2.35 (s,6H), 1.38 (t, J=7.1 Hz, 3H); ES-LCMS m/z 522.2 (M+H).

Example 3N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride, hydrate 1

A suspension ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide(350 mg) in 8.4 mL of acetone:water (9:1, v/v) was stirred for 15 min at23° C. HCl (5M solution in water, 1.0 equivalent) was added and thesample was heated to 40° C. The sample was stirred andtemperature-cycled between 40° C. and 5° C. for 24 h and then stirred at20° C. for 0.5 h. The solids were isolated by vacuum filtration anddried in a vacuum oven at 40° C. for at least 16 hours to give the titlecompound as a crystalline solid.

Alternative Procedure

A slurry ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride, anhydrate (500 mg) in water (12 mL) wastemperature-cycled between 40° C. and 5° C. over 2 d. The sample washarvested by filtration and dried to give the title compound as acrystalline solid.

The X-ray powder diffraction (XRPD) pattern of this material (hydrate 1of the hydrochloric acid salt of the compound of Formula (I)) is shownin FIG. 1 and a summary of the diffraction angles and d-spacings isgiven in Table I below. The XRPD analysis was conducted on aPANanalytical X'Pert Pro Diffractometer on Si zero-background wafers.The acquisition conditions included: Cu K_(α) radiation, generatortension: 45 kV, generator current: 40 mA, step size: 0.02° 2θ.

TABLE I Diff. Angle [°2 θ] d-spacing [Å] 5.9586 14.8328 6.0888 14.515899.013 9.81187 9.2437 9.56744 11.7804 7.51235 11.8888 7.44413 12.14947.28502 13.3294 6.64266 13.4351 6.59061 13.5841 6.51868 13.9743 6.3375214.84 5.96474 14.9038 5.94426 15.337 5.77733 20.4737 4.33798 22.22373.99688 22.2822 3.98652 24.4687 3.63501 24.6344 3.61095 24.9574 3.5649425.0988 3.54517 25.177 3.53434

The Raman spectrum of the title compound was recorded on a Nicolet NXR9650 FT-Raman Spectrometer, at 4 cm⁻¹ resolution with excitation from aNd:YVO4 laser (λ=1064 nm). The Raman spectrum of this material is shownin FIG. 2 with major peaks observed at 455.6, 581.0, 697.7, 773.5,809.3, 951.5, 999.3, 1029.7, 1109.3, 1172.1, 1247.8, 1282.4, 1334.7,1361.6, 1461.3, 1531.6, 1626.3, 1677.7, 2902.7, 2951.6, and 3032.8 cm⁻¹.

The differential scanning calorimetry (DSC) thermogram of the titlecompound was recorded on a TA Instruments Q100 Differential Scanningcalorimeter equipped with an autosampler and a refrigerated coolingsystem under 40 mL/min N₂ purge and is shown in FIG. 3. The experimentswere conducted using a heating rate of 15° C./min in a crimped aluminumpan. The DSC thermogram of hydrate 1 of the hydrochloric acid salt ofthe compound of Formula (I) exhibited a first endotherm with an onsettemperature of about 32° C., a peak temperature about 74° C., andenthalpy of 44.07 J/g, followed by a second endotherm with an onsettemperature of about 158° C., a peak temperature about 173° C., andenthalpy of 55.24 J/g. A person skilled in the art would recognize thatthe onset temperature, peak temperature, and enthalpy of the endothermmay vary depending on the experimental conditions.

The thermogravimetric analysis (TGA) thermogram of the title compoundwas recorded on a TA Instruments Q500 Thermogravimetric Analyzer and isshown in FIG. 4. The experiments were conducted with 40 mL/min N₂ flowand a heating rate of 15° C./min. The TGA thermogram of hydrate 1 of thehydrochloric acid salt of the compound of Formula (I) exhibited twosteps of weight loss events observed prior to the final thermaldecomposition. The first weight loss event takes place in thetemperature range of 30° C. to 100° C. with a weight loss of 1.52%. Thesecond weight loss event takes place in the temperature range of 100° C.to 200° C. with a weight loss of 2.57%.

Example 4N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride, hydrate 2

A suspension ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride, hydrate 1 (39.1 mg) in 1 mL of acetone:water (9:1, v/v)was stirred and temperature-cycled between 40° C. and 5° C. for 3 d. Thefiltrate is then evaporated at room temperature for seven days to givethe title compound as a crystalline solid.

The X-ray powder diffraction (XRPD) pattern of this material (hydrate 2of the hydrochloric acid salt of the compound of Formula (I)) is shownin FIG. 5 and a summary of the diffraction angles and d-spacings isgiven in Table II below. The XRPD analysis was conducted on aPANanalytical X'Pert Pro Diffractometer on Si zero-background wafers.The acquisition conditions included: Cu K_(α) radiation, generatortension: 45 kV, generator current: 40 mA, step size: 0.02° 2θ.

TABLE II Diff. Angle [°2 θ] d-spacing [Å] 6.0237 14.67257 8.9915 9.8352312.0697 7.33295 13.4151 6.60039 13.518 6.55036 14.7032 6.01993 14.76226.00096 14.8596 5.96186 15.2698 5.80263 20.2834 4.37464 20.3721 4.3593922.1624 4.01112 22.2749 3.9878 22.3759 3.97991 24.7082 3.60033 24.77043.60035

The Raman spectrum of the title compound was recorded on a Nicolet NXR9650 FT-Raman Spectrometer, at 4 cm⁻¹ resolution with excitation from aNd:YVO4 laser (λ=1064 nm). The Raman spectrum of this material is shownin FIG. 6 with major peaks observed at 455.3, 588.2, 699.4, 734.2,775.3, 806.8, 884.7, 948.6, 1000.0, 1033.3, 1112.3, 1180.6, 1247.3,1269.2, 1282.5, 1331.8, 1365.7, 1424.7, 1466.3, 1530.0, 1549.9, 1569.7,1627.3, 1683.8, 2901.8, 2946.4, and 3044.2 cm⁻¹.

The differential scanning calorimetry (DSC) thermogram of the titlecompound was recorded on a TA Instruments Q100 Differential Scanningcalorimeter equipped with an autosampler and a refrigerated coolingsystem under 40 mL/min N₂ purge and is shown in FIG. 7. The experimentswere conducted using a heating rate of 15° C./min in a crimped aluminumpan. The DSC thermogram of hydrate 2 of the hydrochloric acid salt ofthe compound of Formula (I) exhibited a first endotherm with an onsettemperature of about 66° C., a peak temperature about 82° C., andenthalpy of 26.177 J/g, followed by a second endotherm with an onsettemperature of about 166° C., a peak temperature about 181° C., andenthalpy of 65.71 J/g. A person skilled in the art would recognize thatthe onset temperature, peak temperature, and enthalpy of the endothermmay vary depending on the experimental conditions.

The thermogravimetric analysis (TGA) thermogram of the title compoundwas recorded on a TA Instruments Q500 Thermogravimetric Analyzer and isshown in FIG. 8. The experiments were conducted with 40 mL/min N₂ flowand a heating rate of 15° C./min. The TGA thermogram of hydrate 2 of thehydrochloric acid salt of the compound of Formula (I) exhibited twosteps of weight loss events observed prior to the final thermaldecomposition. The first weight loss event takes place in thetemperature range of 30° C. to 100° C. with a weight loss of 2.43%. Thesecond weight loss event takes place in the temperature range of 100° C.to 220° C. with a weight loss of 3.29%.

Example 5N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride, anhydrate

A slurry ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehydrochloride, hydrate 1 (500 mg) in 12.5 mL of EtOH was stirred andtemperature-cycled between 40° C. and 5° C. for 1 d. The sample washarvested by filtration and air-dried for a minimum of 1 h to give thetitle compound as a crystalline solid.

The X-ray powder diffraction (XRPD) pattern of this material (theanhydrous hydrochloric acid salt of the compound of Formula (I)) isshown in FIG. 9 and a summary of the diffraction angles and d-spacingsis given in Table III below. The XRPD analysis was conducted on aPANanalytical X'Pert Pro Diffractometer on Si zero-background wafers.The acquisition conditions included: Cu K_(α) radiation, generatortension: 45 kV, generator current: 40 mA, step size: 0.02° 2θ.

TABLE III Diff. Angle [°2 θ] d-spacing [Å] 8.2804 10.67826 8.358510.57856 10.6756 8.28718 11.2515 7.86431 15.529 5.70635 16.0376 5.5265219.9806 4.44392 20.3778 4.35818 20.7946 4.27176 22.6327 3.92883 23.22783.82633 23.3175 3.81497 23.6252 3.76597 24.5698 3.62028 24.6174 3.6223824.902 3.57274 25.2799 3.52018 25.8512 3.44367 26.8957 3.31225 27.25723.26914 27.3858 3.25408 28.0514 3.17837 28.1463 3.16786 28.2384 3.16559

The Raman spectrum of the title compound was recorded on a Nicolet NXR9650 FT-Raman Spectrometer, at 4 cm⁻¹ resolution with excitation from aNd:YVO4 laser (λ=1064 nm). The Raman spectrum of this material is shownin FIG. 10 with major peaks observed at 418.4, 453.5, 575.3, 635.8,699.3, 771.0, 782.0, 805.0, 863.9, 893.8, 940.5, 974.2, 997.9, 1058.1,1115.8, 1190.2, 1245.9, 1272.8, 1299.2, 1328.5, 1355.7, 1406.8, 1433.1,1462.1, 1489.0, 1511.0, 1546.3, 1562.1, 1614.1, 1625.9, 1666.7, 1695.0,2921.9, 2950.2, 2986.2, 3036.1, 3075.0, and 3095.0 cm⁻¹.

The differential scanning calorimetry (DSC) thermogram of the titlecompound was recorded on a TA Instruments Q100 Differential Scanningcalorimeter equipped with an autosampler and a refrigerated coolingsystem under 40 mL/min N₂ purge and is shown in FIG. 11. The experimentswere conducted using a heating rate of 15° C./min in a crimped aluminumpan. The DSC thermogram of the anhydrous hydrochloric acid salt of thecompound of Formula (I) exhibited a sharp endotherm with an onsettemperature of 256.44° C., a peak temperature about 258.48° C., andenthalpy of 128.0 J/g. A person skilled in the art would recognize thatthe onset temperature, peak temperature, and enthalpy of the endothermmay vary depending on the experimental conditions.

The thermogravimetric analysis (TGA) thermogram of the title compoundwas recorded on a TA Instruments Q500 Thermogravimetric Analyzer and isshown in FIG. 12. The experiments were conducted with 40 mL/min N₂ flowand a heating rate of 15° C./min. The TGA thermogram of the anhydroushydrochloric acid salt of the compound of Formula (I) exhibitednegligible weight loss in the temperature range of 25° C. to 150° C. anda thermal decomposition onset temperature of 267.33° C.

Example 6N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate

A suspension ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide(353.1 mg) in 8.4 mL of acetone was stirred for 15 min at 23° C.L-aspartic acid (99% purity, 85.1 mg, powder, 1.0 equivalent) was addedand the sample was heated to 40° C. and then seeded withN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamideaspartate. The sample was stirred and temperature-cycled between 40° C.and 5° C. for 6 h and then stirred at 20° C. for 0.5 h. The solids wereisolated by vacuum filtration and dried in a vacuum oven at 40° C. forat least 16 hours to give the title compound as a crystalline solid. ¹HNMR analysis (500 MHz, DMSO-d₆) indicated 1:1 acid:free basestoichiometry.

Seed Preparation

A suspension ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide(25 mg) in 0.6 mL of acetone was stirred for 30 min at 23° C. L-asparticacid (6.1 mg) was added and the sample was heated to 40° C. The samplewas stirred and temperature-cycled between 40° C. and 5° C. for 48 h,then stirred at 20° C. for 24 h, and then at 4° C. for 24 h. The solidswere isolated by vacuum filtration and dried to give the title compoundas a crystalline solid.

The X-ray powder diffraction (XRPD) pattern of this material (theaspartic acid salt of the compound of Formula (I)) is shown in FIG. 13and a summary of the diffraction angles and d-spacings is given in TableIV below. The XRPD analysis was conducted on a PANanalytical X'Pert ProDiffractometer on Si zero-background wafers. The acquisition conditionsincluded: Cu K_(α) radiation, generator tension: 45 kV, generatorcurrent: 40 mA, step size: 0.02° 2θ.

TABLE IV Diff. Angle [°2 θ] d-spacing [Å] 6.7436 13.10777 6.960212.70047 7.6442 11.5654 11.7912 7.50553 13.9213 6.36149 14.7678 5.9937715.5507 5.69372 15.6273 5.66596 15.7559 5.62001 16.241 5.45324 17.72335.00036 18.4499 4.80504 18.7127 4.73814 19.0754 4.64886 19.1568 4.6292919.2284 4.62368 20.0643 4.42191 20.6095 4.30614 20.9589 4.23513 21.06454.21415 21.1555 4.20665 21.704 4.09141 22.0986 4.01922 22.8012 3.8969423.0234 3.85984 23.1372 3.84111 23.2967 3.81516 23.694 3.75209 23.76743.74996 24.9715 3.56296 25.0549 3.55129 25.4173 3.50146 25.5014 3.4901125.6884 3.46513 25.7417 3.45806 26.1759 3.40168 27.399 3.25254 28.21663.16013 31.1595 2.86805 35.8675 2.50164 35.9564 2.50187

The Raman spectrum of the title compound was recorded on a Nicolet NXR9650 FT-Raman Spectrometer, at 4 cm⁻¹ resolution with excitation from aNd:YVO4 laser (λ=1064 nm). The Raman spectrum of this material is shownin FIG. 14 with major peaks observed at 452.2, 573.8, 690.5, 770.0,806.8, 999.6, 1036.5, 1106.3, 1162.4, 1236.9, 1274.1, 1332.1, 1363.6,1470.7, 1487.4, 1529.5, 1627.1, 1704.6, 2917.9, and 3072.7 cm⁻¹.

The differential scanning calorimetry (DSC) thermogram of the titlecompound was recorded on a TA Instruments Q100 Differential Scanningcalorimeter equipped with an autosampler and a refrigerated coolingsystem under 40 mL/min N₂ purge and is shown in FIG. 15. The experimentswere conducted using a heating rate of 15° C./min in a crimped aluminumpan. The DSC thermogram of the aspartic acid salt of the compound ofFormula (I) exhibited a sharp endotherm with an onset temperature of220.62° C., a peak temperature about 223.04° C., and enthalpy of 75.76J/g. A person skilled in the art would recognize that the onsettemperature, peak temperature, and enthalpy of the endotherm may varydepending on the experimental conditions.

The thermogravimetric analysis (TGA) thermogram of the title compoundwas recorded on a TA Instruments Q500 Thermogravimetric Analyzer and isshown in FIG. 16. The experiments were conducted with 40 mL/min N₂ flowand a heating rate of 15° C./min. The TGA thermogram of the asparticacid salt of the compound of Formula (I) exhibited negligible weightloss in the temperature range of 25° C. to 150° C. and a thermaldecomposition onset temperature of 240.39° C.

Example 7N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate

A suspension ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide(350 mg) in 8.4 mL of acetone was stirred for 15 min at 23° C. Hippuricacid (98% purity, 117.3 mg, powder, 1.0 equivalent) was added and thesample was heated to 40° C. and then seeded withN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidehippurate. The sample was stirred and temperature-cycled between 40° C.and 5° C. for 6 h and then stirred at 20° C. for 0.5 h. The solids wereisolated by vacuum filtration and dried in a vacuum oven at 40° C. forat least 16 hours to give the title compound as a crystalline solid. ¹HNMR analysis (500 MHz, DMSO-d₆) indicated 1:1 acid:free basestoichiometry.

Seed Preparation

A suspension ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide(25 mg) in 0.6 mL of acetone was stirred for 30 min at 23° C. Hippuricacid (8.4 mg) was added and the sample was heated to 40° C. The samplewas stirred and temperature-cycled between 40° C. and 5° C. for 48 h,then stirred at 20° C. for 24 h, and then at 4° C. for 24 h. The solidswere isolated by vacuum filtration and dried to give the title compoundas a crystalline solid.

The X-ray powder diffraction (XRPD) pattern of this material (thehippuric acid salt of the compound of Formula (I)) is shown in FIG. 17and a summary of the diffraction angles and d-spacings is given in TableV below. The XRPD analysis was conducted on a PANanalytical X'Pert ProDiffractometer on Si zero-background wafers. The acquisition conditionsincluded: Cu K_(α) radiation, generator tension: 45 kV, generatorcurrent: 40 mA, step size: 0.02° 2θ.

TABLE V Diff. Angle [°2 θ] d-spacing [Å] 6.1274 14.42462 9.2336 9.5778810.068 8.78592 12.0304 7.35682 12.336 7.17522 12.625 7.01165 13.77926.42678 14.0043 6.32399 17.2054 5.15396 17.337 5.11513 18.0894 4.9040418.3887 4.82487 18.9044 4.6944 19.022 4.66563 19.1238 4.64104 19.59974.52941 20.619 4.30774 21.1226 4.20269 21.2004 4.19785 21.5393 4.1223121.7178 4.08883 22.4194 3.96243 22.8549 3.88791 23.4931 3.78371 23.63453.7614 24.9064 3.57212 27.2014 3.27573 27.3599 3.2571 27.4394 3.2559227.6333 3.2255 27.96 3.18854

The Raman spectrum of the title compound was recorded on a Nicolet NXR9650 FT-Raman Spectrometer, at 4 cm⁻¹ resolution with excitation from aNd:YVO4 laser (λ=1064 nm). The Raman spectrum of this material is shownin FIG. 18 with major peaks observed at 556.6, 618.1, 694.0, 771.4,809.0, 916.5, 997.2, 1041.9, 1108.4, 1235.8, 1272.2, 1335.1, 1366.2,1466.6, 1537.0, 1575.4, 1601.4, 1630.2, 1695.0, 2943.8, and 3071.4 cm⁻¹.

The differential scanning calorimetry (DSC) thermogram of the titlecompound was recorded on a TA Instruments Q100 Differential Scanningcalorimeter equipped with an autosampler and a refrigerated coolingsystem under 40 mL/min N₂ purge and is shown in FIG. 19. The experimentswere conducted using a heating rate of 15° C./min in a crimped aluminumpan. The DSC thermogram of the hippuric acid salt of the compound ofFormula (I) exhibited a sharp endotherm with an onset temperature of232.10° C., a peak temperature about 233.32° C., and enthalpy of 123.1J/g. A person skilled in the art would recognize that the onsettemperature, peak temperature, and enthalpy of the endotherm may varydepending on the experimental conditions.

The thermogravimetric analysis (TGA) thermogram of the title compoundwas recorded on a TA Instruments Q500 Thermogravimetric Analyzer and isshown in FIG. 20. The experiments were conducted with 40 mL/min N₂ flowand a heating rate of 15° C./min. The TGA thermogram of the hippuricacid salt of the compound of Formula (I) exhibited negligible weightloss in the temperature range of 25° C. to 150° C. and a thermaldecomposition onset temperature of 245.70° C.

Example 8N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate

A suspension ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide(352 mg) in 8.4 mL of acetone was stirred for 15 min at 23° C.Phosphoric acid (3.0 M solution in water, 1.0 equivalent) was added andthe sample was heated to 40° C. and then seeded withN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamidephosphate. The sample was stirred and temperature-cycled between 40° C.and 5° C. for 6 h and then stirred at 20° C. for 0.5 h. The solids wereisolated by vacuum filtration and dried in a vacuum oven at 40° C. forat least 16 hours to give the title compound as a crystalline solid.Inductively coupled plasma atomic emission spectroscopy indicated 1:1acid:free base stoichiometry.

Seed Preparation

A suspension ofN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide(25 mg) in 0.6 mL of acetone was stirred for 30 min at 23° C. Phosphoricacid (3.0 M solution in water, 1.0 equivalent) was added and the samplewas heated to 40° C. The sample was stirred and temperature-cycledbetween 40° C. and 5° C. for 48 h, then stirred at 20° C. for 24 h, andthen at 4° C. for 24 h. The solids were isolated by vacuum filtrationand dried to give the title compound as a crystalline solid.

The X-ray powder diffraction (XRPD) pattern of this material (thephosphoric acid salt of the compound of Formula (I)) is shown in FIG. 21and a summary of the diffraction angles and d-spacings is given in TableVI below. The XRPD analysis was conducted on a PANanalytical X'Pert ProDiffractometer on Si zero-background wafers. The acquisition conditionsincluded: Cu K_(α) radiation, generator tension: 45 kV, generatorcurrent: 40 mA, step size: 0.02° 2θ.

TABLE VI Diff. Angle [°2 θ] d-spacing [Å] 5.51 16.03945 5.605 15.767647.6688 11.52832 9.9043 8.93072 11.1846 7.91119 15.352 5.77173 15.95545.5548 16.8019 5.27679 18.1765 4.88073 19.8809 4.46227 20.3291 4.364923.8956 3.72088 24.1905 3.67619 24.4455 3.63841 26.5298 3.3571 26.72773.3327 27.0256 3.29663 28.6856 3.10952

The Raman spectrum of the title compound was recorded on a Nicolet NXR9650 FT-Raman Spectrometer, at 4 cm⁻¹ resolution with excitation from aNd:YVO4 laser (λ=1064 nm). The Raman spectrum of this material is shownin FIG. 22 with major peaks observed at 460.6, 484.5, 529.3, 577.0,637.8, 696.3, 731.9, 773.1, 786.2, 805.7, 866.9, 888.7, 1002.3, 1035.6,1187.1, 1242.7, 1275.9, 1296.0, 1326.2, 1357.6, 1375.1, 1441.7, 1466.0,1510.1, 1531.6, 1580.3, 1624.5, 1698.2, 2936.4, 2964.1, and 3068.6 cm⁻¹.

The differential scanning calorimetry (DSC) thermogram of the titlecompound was recorded on a TA Instruments Q100 Differential Scanningcalorimeter equipped with an autosampler and a refrigerated coolingsystem under 40 mL/min N₂ purge and is shown in FIG. 23. The experimentswere conducted using a heating rate of 15° C./min in a crimped aluminumpan. The DSC thermogram of the phosphoric acid salt of the compound ofFormula (I) exhibited a sharp endotherm with an onset temperature of235.69° C., a peak temperature about 242.24° C., and enthalpy of 135.3J/g. A person skilled in the art would recognize that the onsettemperature, peak temperature, and enthalpy of the endotherm may varydepending on the experimental conditions.

The thermogravimetric analysis (TGA) thermogram of the title compoundwas recorded on a TA Instruments Q500 Thermogravimetric Analyzer and isshown in FIG. 24. The experiments were conducted with 40 mL/min N₂ flowand a heating rate of 15° C./min. The TGA thermogram of the phosphoricacid salt of the compound of Formula (I) exhibited negligible weightloss in the temperature range of 25° C. to 150° C. and a thermaldecomposition onset temperature of 237.30° C.

Biological Assays

The compounds of the present invention were tested for RET kinaseinhibitory activity in a RET kinase enzyme assay, a cell-basedmechanistic assay and a cell-based proliferation assay.

RET Kinase Enzymatic Assay

Human RET kinase cytoplasmic domain (amino acids 658-1114 of accessionnumber NP_000314.1) was expressed as an N-terminal GST-fusion proteinusing a baculovirus expression system. GST-RET was purified usingglutathione sepharose chromatography. The RET kinase enzymatic assay wasperformed in a total volume of 10 uL with increasing concentrations ofRET kinase inhibitor as a singlet in a 384 well format as follows: RETinhibitor compound plates are prepared by adding 100 nL of RET inhibitorat different concentrations to a 384-well plate. 5 μL/well of a 2×enzyme mix (50 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonicacid); 1 mM CHAPS(3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate); 0.1 mg/mLBSA (bovine serum albumin); 1 mM DTT (dithiothreitol); 0.2 nM RETkinase) was added to the 384-well plate and incubated for 30 minutes at23° C. 5 μL/well of a 2× substrate mix (50 mM HEPES; 1 mM CHAPS; 0.1mg/mL BSA; 20 μM adenosine triphosphate; 20 mM MgCl₂ and 1 μMbiotinylated peptide substrate) was added and incubated for 1 hour at23° C. 10 μL/well of 2× stop/detection mix (50 mM HEPES; 0.1% BSA; 800mM Potassium Fluoride; 50 mM EDTA (Ethylenediaminetetraacetic acid);200× dilution of Europium Cryptate labeled anti-phosphotyrosineantibody; 62.5 nM Streptavidin-XL665) incubated for 1 hour at 23° C. andread on a Homogenous Time-Resolved Fluorescence reader. IC₅₀s werefitted using GraphPad Prism to a sigmoidal dose response.

RET Kinase Cell-Based Mechanistic Assay

The potency of the compounds of the invention were tested for itsability to inhibit constitutive RET kinase phosphorylation in cell-basedassay. TT cells (ATCC CRL-1803), a medullary thyroid cancer cell linewith constitutively activated RET kinase, were maintained in 150 cm²dishes in F12 Kaighn's medium, 10% fetal bovine serum, 1× Glutamax, 1×non-essential amino acids, 1× Pen/Strep antibiotics at 37° C. in 5%carbon dioxide. 1.0E5 TT cells/well were plated in a 96-well cellculture plate and allowed to adhere overnight. TT cells were treatedwith different concentrations of RET inhibitor compounds for 2 h at 37°C. in 5% carbon dioxide, washed with ice cold PBS (phosphate bufferedsaline) and lysed by adding 200 μL of 25 mM Tris HCl pH 7.5; 2 mM EDTA;150 mM NaCl; 1% sodium deoxycholate; 1% Triton X-100; 50 mM sodium betaglycerophosphate; 1 mM sodium orthovanadate; 1× phosphatase inhibitorcocktail #2 (Sigma # P5726); 1× phosphatase inhibitor cocktail #3 (Sigma# P0044) and 1× complete mini EDTA free protease inhibitor cocktail(Roche #4693159001), incubation at −80° C. for 10 minutes and thawed onice. 100 μL of TT cell lysate was added to a 96-well plate overnight at4° C. that had been coated overnight at 4° C. with 1:1,000 dilution of arabbit anti-RET antibody (Cell Signaling #7032) blocked with 1×PBS;0.05% Tween-20; 1% bovine serum albumin. Plates were washed 4× with 200μL of 1×PBS; 0.05% Tween-20 and then 100 μL of a 1:1,000 dilution of ananti-phosphotyrosine detection antibody (Cell Signaling #7034) was addedand incubated for 1 hour at 37° C. Plates were washed 4× with 200 μL of1×PBS; 0.05% Tween-20 and then 100 μL of a 1:1,000 dilution of ananti-mouse immunoglobulin horse radish peroxidase conjugate antibody(Cell Signaling #7034) was added and incubated for 30 minutes at 37° C.Plates were washed 4× with 200 μL of 1×PBS; 0.05% Tween-20, 100 μL ofTMB (3,3′,5,5″-tetramethylbenzidine) substrate (Cell Signaling #7004)was added, incubated for 10 minutes at 37° C., 100 μL of Stop solution(Cell Signaling #7002) was added and absorbance read on aspectrophotometer at 450 nm. IC₅₀s were fitted using GraphPad Prism to asigmoidal dose response.

RET Kinase Cell-Based Proliferation Assay

The potency of the compounds of the invention were tested for itsability to inhibit cell proliferation and cell viability. TT cells (ATCCCRL-1803), a medullary thyroid cancer cell line with constitutivelyactivated RET kinase, were maintained in 150 cm² dishes in F12 Kaighn'smedium, 10% fetal bovine serum, 1× Glutamax, 1× non-essential aminoacids, 1× Pen/Strep antibiotics at 37° C. in 5% carbon dioxide. 6.0E3 TTcells/well in 50 μL of media were added to a 96-well cell culture plateand allowed to adhere overnight. 50 μL of serially diluted RET inhibitorcompounds were added to 96-well plate containing cultured TT cells andincubated at at 37° C. in 5% carbon dioxide for eight days. 50 μL ofCellTiter-Glo (Promega # G-7573) was added, contents mixed for 1 minuteon shaker followed by 10 minutes in the dark at 23° C. and theluminescence read by EnVision (PerkinElmer). IC₅₀s were fitted usingGraphPad Prism to a sigmoidal dose response.

Biological Data

Exemplified compounds of the present invention were tested in the RETassays described above and were found to be inhibitors of RET withIC₅₀<10 μM. Data for specific examples tested are listed below in Table1 as follows: +=10 μM>IC₅₀>100 nM; ++=100 nM≥IC₅₀>10 nM; +++=IC₅₀≤10 nM.

TABLE 1 Human RET kinase Human RET kinase Exam- Human RET kinasecell-based cell-based ple # enzymatic IC₅₀ mechanistic IC₅₀proliferation IC₅₀ 1 +++ ++ ++ 2 +++ ++ ++In Vivo Colonic Hypersensitivity Model

The efficacy of RET kinase inhibitor compounds can be evaluated in an invivo model of colonic hypersensitivity (Hoffman, J. M., et al.,Gastroenterology, 2012, 142:844-854).

The invention claimed is:
 1. A method of treating a disease or conditioncomprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound which isN-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(trifluoromethyl)benzamide,represented by Formula (I):

or a pharmaceutically acceptable salt thereof; wherein said disease orcondition is selected from irritable bowel syndrome (IBS), painassociated with IBS, functional bloating, functional constipation,functional diarrhea, unspecified functional bowel disorder, functionalabdominal pain syndrome, chronic idiopathic constipation, functionalesophageal disorders, functional gastroduodenal disorders, functionalanorectal pain, inflammatory bowel disease, and proliferative diseases.2. A method according to claim 1 wherein the compound is in the form ofa pharmaceutically acceptable salt selected from the group consisting ofthe hydrochloric acid salt, aspartic acid salt, hippuric acid salt, andphosphoric acid salt.
 3. A method according to claim 1 wherein thecompound is in the form of a hydrochloric acid salt.
 4. A methodaccording to claim 1 wherein the compound or pharmaceutically acceptablesalt thereof is crystalline.
 5. A method according to claim 1 whereinthe disease or condition is IBS.
 6. A method according to claim 5wherein the IBS is diarrhea-predominant, constipation-predominant oralternating stool pattern.
 7. A method according to claim 5 wherein theIBS is diarrhea-predominant.
 8. A method according to claim 1 whereinthe disease or condition is pain associated with IBS.
 9. A methodaccording to claim 1 wherein the disease or condition is functionalbloating.
 10. A method according to claim 1 wherein the disease orcondition is functional constipation.
 11. A method according to claim 1wherein the disease or condition is functional diarrhea.
 12. A methodaccording to claim 1 wherein the disease or condition is inflammatorybowel disease.
 13. A method according to claim 1 wherein the disease orcondition is a proliferative disorder selected from non-small cell lungcancer, hepatocellular carcinoma, colorectal cancer, medullary thyroidcancer, follicular thyroid cancer, anaplastic thyroid cancer, papillarythyroid cancer, brain tumors, peritoneal cavity cancer, solid tumors,other lung cancer, head and neck cancer, gliomas, neuroblastomas, VonHippel-Lindau Syndrome and kidney tumors, breast cancer, fallopian tubecancer, ovarian cancer, transitional cell cancer, prostate cancer,cancer of the esophagus and gastroesophageal junction, biliary cancerand adenocarcinoma or a combination thereof.
 14. A method according toclaim 1 wherein the subject is a mammal.
 15. A method according to claim1 wherein the subject is a human.